Derivatives of antifungal substance BE-31405 and process for producing the same

ABSTRACT

The present invention relates to a compound represented by general formula (I) or a pharmaceutically acceptable salt or ester thereof: ##STR1## wherein each of R 1  and R 2  is independently a hydrogen atom, a C 1  -C 16  alkyl group, a C 2  -C 10  alkenyl group, a C 3  -C 6  alkynyl group, a C 6  -C 12  aryl group, a C 7  -C 15  aralkyl group or a heterocyclic group which is not substituted, a C 1  -C 16  alkyl group, a C 2  -C 10  alkenyl group, a C 3  -C 6  alkynyl group, a C 6  -C 12  aryl group, a C 7  -C 15  aralkyl group or a heterocyclic group which has substituent(s), or a group represented by --Y--R 3  (provided that when R 2  is a hydrogen atom, R 1  is not a methyl group or an acetyl group; when R 2  is an acetyl group, R 1  is not a methyl group), an antifungal agent containing it as an active ingredient and an antifungal composition containing it and an azole type antifungal agent.

This application is a 371 of PCT/JP98/02218 filed on May 20, 1998.

TECHNICAL FIELD

The present invention is useful in the pharmaceutical field. Morespecifically, the present invention relates to a novel antifungal agent.

BACKGROUND ART

In the field of antifungal agents, a number of compounds have alreadycome into practical use as pharmaceuticals. However, their effects onvarious harmful strains are not always satisfactory, and emergence ofstrains resistant to these pharmaceuticals, especially to azole typeantifungal agents in wide use, has become a serious clinical problem.Therefore, development of pharmaceuticals effective against theseharmful strains and resistant strains is demanded.

JP-A-6-157582, J. Antibiotics, vol. 48, pp.1171-1172 (1995) and Nat.Prod. Lett., vol. 7, pp.309-316 (1995) disclose structural analogues ofthe compounds of the present invention which show excellent antifungalaction, but neither disclose nor suggest anything specific about thecompounds of the present invention.

Further, the present inventors have recently found that compoundscontaining the compounds disclosed by the above-mentioned references andazole type antifungal agents have excellent antifungal action(PCT/JP96/00353).

However, development of even more excellent antifungal agents is stilldemanded.

DISCLOSURE OF THE INVENTION

The object of the present invention is to provide novel antifungalagents which meet the above-mentioned demand. Namely, the problem thatthe present invention is to solve is to provide agents which showantifungal action against various harmful strains and resistant strainsagainst which conventional antifungal agents are not satisfactorilyeffective.

The present inventors have conducted extensive research to solve theabove-mentioned problem, and as a result, have found that the compoundsrepresented by general formula (I) and pharmaceutically acceptable saltsor esters thereof have excellent antifungal activities, and the presentinvention has been accomplished.

Namely, the present invention provides a compound represented by generalformula (I) or a pharmaceutically acceptable salt or ester thereof:##STR2## wherein each of R¹ and R² is independently a hydrogen atom, aC₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, aC₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic groupwhich is not substituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup or a heterocyclic group which has one to five substituentsselected from the group consisting of halogen atoms, cyano groups,hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxygroups, amino groups, mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonyl groups,aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxygroups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or a grouprepresented by --Y--R³ ; Y is a carbonyl group, a thiocarbonyl group ora sulfonyl group; and R³ is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which is not substituted, ora C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkylgroup, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, aC₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or a heterocyclicgroup which has one to four substituents selected from the groupconsisting of halogen atoms, cyano groups, hydroxyl groups, amino groupsand carboxyl groups, and hydroxyl groups, amino groups and carboxylgroups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkyl group, ahydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, a carboxy-C₁-C₁₆ alkyl group or a protecting group (provided that when R² is ahydrogen atom, R¹ is not a methyl group or an acetyl group; when R² isan acetyl group, R¹ is not a methyl group) and an antifungal agentcontaining it as an active ingredient.

The present invention also provides an antifungal composition containinga compound represented by general formula (I) or a pharmaceuticallyacceptable salt or ester thereof and an azole type antifungal agent asactive ingredients.

The symbols and terms used in the specification will be explained.

A C₁ -C₁₆ alkyl group means a linear or branched alkyl group having acarbon number of 1 to 16, such as a methyl group, an ethyl group, apropyl group, an isopropyl group, a butyl group, an isobutyl group, asec-butyl group, a tert-butyl group, a pentyl group, an isopentyl group,a neopentyl group, a hexyl group, a decyl group, a dodecyl group or ahexadecyl group.

A C₂ -C₁₀ alkenyl group means a linear or branched alkenyl group whichhas one to five double bonds and a carbon number of 2 to 10, such as apropenyl group, a 2-butenyl group, a 3-butenyl group, a 3-pentenylgroup, a 4-hexenyl group or a 1,3-hexadienyl group.

A C₃ -C₆ alkynyl group means a linear or branched alkynyl group whichhas one to three triple bonds and a carbon number of 3 to 6, such as apropynyl group, a 2-butynyl group, a 3-butynyl group, a 3-pentynylgroup, a 4-hexynyl group or a 1-decynyl group.

A C₆ -C₁₂ aryl group means a monocyclic or polycyclic aryl group havinga carbon number of 6 to 12, such as a phenyl group, a naphthyl group ora tetrahydronaphthyl group.

A C₆ -C₁₂ aryloxy group means an aryloxy group having theabove-mentioned C₆ -C₁₂ aryl group, such as a phenyloxy group, anaphthyloxy group or a tetrahydronaphthyloxy group.

A halogen atom means a fluorine atom, a chlorine atom, a bromine atom oran iodine atom.

A C₁ -C₁₆ alkyloxy group means an alkyloxy group having theabove-mentioned C₁ -C₁₆ alkyl group, such as a methyloxy group, anethyloxy group, a propyloxy group, an isopropyloxy group, a butyloxygroup, an isobutyloxy group, a sec-butyloxy group, a tert-butyloxygroup, a pentyloxy group, an isopentyloxy group, a neopentyloxy group, ahexyloxy group, a decyloxy group, a dodecyloxy group, a hexadecyloxygroup or a cetyloxy group.

A C₁ -C₁₆ alkyloxycarbonyl group means an alkyloxycarbonyl group havingthe above-mentioned C₁ -C₁₆ alkyl group, such as a methyloxycarbonylgroup, an ethyloxycarbonyl group, a propyloxycarbonyl group, anisopropyloxycarbonyl group, a butyloxycarbonyl group, anisobutyloxycarbonyl group, a sec-butyloxycarbonyl group, atert-butyloxycarbonyl group, a pentyloxycarbonyl group, anisopentyloxycarbonyl group, a neopentyloxycarbonyl group, ahexyloxycarbonyl group, a decyloxycarbonyl group, a dodecyloxycarbonylgroup or a hexadecyloxycarbonyl group.

A C₁ -C₁₆ alkylcarbonyloxy group means an alkylcarbonyloxy group havingthe above-mentioned C₁ -C₁₆ alkyl group, such as a methylcarbonyloxygroup, an ethylcarbonyloxy group, a propylcarbonyloxy group, anisopropylcarbonyloxy group, a butylcarbonyloxy group, anisobutylcarbonyloxy group, a sec-butylcarbonyloxy group, atert-butylcarbonyloxy group, a pentylcarbonyloxy group, anisopentylcarbonyloxy group, a neopentylcarbonyloxy group, ahexylcarbonyloxy group, a decylcarbonyloxy group, a dodecylcarbonyloxygroup, a hexadecylcarbonyloxy group or a palmitoyloxy group.

A mono-C₁ -C₁₆ alkylamino group means an amino group mono-substitutedwith the above-mentioned C₁ -C₁₆ alkyl group, such as a methylaminogroup, an ethylamino group, a propylamino group, an isopropylaminogroup, a butylamino group, an isobutylamino group, a sec-butylaminogroup, a tert-butylamino group, a pentylamino group, an isopentylaminogroup, a neopentylamino group or a hexylamino group.

A di-C₁ -C₁₆ alkylamino group means an amino group di-substituted withthe above-mentioned C₁ -C₁₆ alkyl groups, such as a dimethylamino group,an ethylmethylamino group, a diethylamino group, an ethylpropylaminogroup, a dipropylamino group, a butylmethylamino group, a dibutylaminogroup, butylethylamino group, a methylpentylamino group, ahexylmethylamino group or an ethylhexylamino group.

A C₃ -C₆ cycloalkyl group means a cycloalkyl group having a carbonnumber of 3 to 6, such as a cyclopropyl group, a cyclobutyl group, acyclopentyl group or a cyclohexyl group.

A C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group means the above-mentioned C₁-C₁₆ alkyl group substituted with the above-mentioned C₃ -C₆ cycloalkylgroup, such as a cyclopropylmethyl group, a cyclobutylmethyl group, acyclopentylmethyl group, a cyclohexylmethyl group, a cyclopropylethylgroup, a cyclobutylethyl group, a cyclopentylethyl group, acyclohexylethyl group, a 3-cyclohexylpropyl group, a 3-cyclopentylpropylgroup, a 4-cyclohexylbutyl group or a 4-cyclopentylbutyl group, andpreferably has a total carbon number of 4 to 10.

A C₇ -C₁₅ aralkyl group means the above-mentioned C₁ -C₁₆ alkyl groupsubstituted with the above-mentioned C₆ -C₁₂ aryl group which has acarbon number of 7 to 15, such as a benzyl group, a phenethyl group, aphenylpropyl group, a phenybutyl group, a phenylpentyl group, anaphthylmethyl group or a naphthylethyl group.

A C₇ -C₁₅ aralkyloxy group means an aralkyloxy group having theabove-mentioned C₇ -C₁₅ aralkyl group, such as a benzyloxy group, aphenethyloxy group, a phenylpropyloxy group, a phenylbutyloxy group, aphenylpentyloxy group, a naphthylmethyloxy group or a naphthylethyloxygroup.

A heterocyclic group means an aromatic or non-aromatic 5 to 7-memberedmonocyclic heterocyclic group having one to four hetero atoms selectedfrom the group consisting of nitrogen atoms, oxygen atoms and sulfuratoms or a condensed heterocyclic group having such a monocyclicheterocyclic group fused with the above-mentioned C₃ -C₆ cycloalkylgroup, the above-mentioned C₆ -C₁₂ aryl group or another identical ordifferent monocyclic heterocyclic group, such as a pyrrolyl group, afuryl group, a thienyl group, an oxazolyl group, an isoxazolyl group, athiazolyl group, an isothiazolyl group, an imidazolyl group, a pyrazolylgroup, an oxadiazoly group, a thiadiazoly group, a triazolyl group, atetrazolyl group, a furazanyl group, a pyridyl group, a pyridazinylgroup, a pyrimidinyl group, a pyrazinyl group, a triazinyl group, adihydrothienyl group, a tetrahydrothienyl group, a pyrrolinyl group, apyrrolidinyl group, an imidazolidinyl group, an imidazolinyl group, apiperidinyl group, a piperazinyl group, an oxazolinyl group, anisoxazolinyl group, an isoxazolidinyl group, a thiazolinyl group, athiazolidinyl group, an isothiazolinyl group, an isothiazolidinyl group,a 1,2-dithiolanyl group, a 1,3-dithiolanyl group, a 1,2-dithiolyl group,a 1,3-dithiolyl group, a dihydrothiopyranyl group, atetrahydrothiopyranyl group, a 1,4-dithianyl group, a 1,4-dithiinylgroup, a 1,4-oxathiinyl group, a thiomorpholinyl group, a morpholinylgroup, an indolyl group, an isoindolyl group, a quionolyl group, anisoquinolyl group, a quinolizinyl group, a cinnolinyl group, aquinoxalinyl group, a phthalazinyl group, a pteridinyl group, a purinylgroup, a carbazolyl group, an acridinyl group, a phenazinyl group, abenzofuryl group, a chromanyl group, an isochromanyl group, a xanthenylgroup, a benzoxazolyl group, an imidazothiazolyl group, athieno[2,3-b]thienyl group or a 1,4-dithianaphthyl group.

A C₇ -C₁₅ aralkylamino group means an aralkylamino group having theabove-mentioned C₇ -C₁₅ aralkyl group, such as a benzylamino group, aphenethylamino group, a phenylpropylamino group, a phenylbutylaminogroup, a phenylpentylamino group, a naphthylmethylamino group or anaphthylethylamino group.

A halo-C₁ -C₁₆ alkyl group means the above-mentioned C₁ -C₁₆ alkyl groupsubstituted with one to three halogen atoms described above, such as afluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a1-fluoroethyl group, a 2-fluoroethyl group, a 1,2-difluoroethyl group, achloromethyl group, a dichloromethyl group, a 1-chloroethyl group, a2-chloroethyl group, a 1,2-dichloroethyl group, a bromomethyl group, adibromomethyl group, a 1-bromoethyl group, a 2-bromoethyl group or a1,2-dibromoethyl group.

A hydroxy-C₁ -C₁₆ alkyl group means the above-mentioned C₁ -C₁₆ alkylgroup substituted with one to three hydroxyl groups, such as ahydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxyethyl group, a1,2-dihydroxyethyl group, a 1-hydroxypropyl group, a 2-hydroxypropylgroup or a 3-hydroxypropyl group.

An amino-C₁ -C₁₆ alkyl group means the above-mentioned C₁ -C₁₆ alkylgroup substituted with one to three amino groups, such as an aminomethylgroup, a 1-aminoethyl group, a 2-aminoethyl group, a 1,2-diaminoethylgroup, a 1-aminopropyl group, a 2-aminopropyl group or a 3-aminopropylgroup.

A carboxy-C₁ -C₁₆ alkyl group means the above-mentioned C₁ -C₁₆ alkylgroup substituted with one to three carboxyl groups, such as acarboxymethyl group, a 1-carboxyethyl group, a 2-carboxyethyl group, a1,2-dicarboxyethyl group, a 1-carboxypropyl group, a 2-carboxypropylgroup or a 3-carboxypropyl group.

A protecting group for a hydroxyl group is a lower alkylsilyl group suchas a trimethylsilyl group or a tert-butyldimethylsilyl group; a loweralkoxymethyl group such as a methoxymethyl group or a2-methoxyethoxymethyl group; a tetrahydropyranyl group; an aralkyl groupsuch as a benzyl group, a p-methoxybenzyl group, a p-nitrobenzyl groupor a trityl group; or an acyl group such as a formyl group or an acetylgroup, preferably a methoxymethyl group, a tetrahydropyranyl group, atrityl group, a tert-butyldimethylsilyl group or an acetyl group.

A protecting group for an amino group is aralkylidene group such as abenzylidene group, a p-chlorobenzylidene group or a p-nitrobenzylidenegroup; an aralkyl group such as a benzyl group, a p-methoxybenzyl group,a p-nitrobenzyl group, a benzhydryl group or a trityl group; a loweralkanoyl group such as formyl group, an acetyl group, a propionyl group,a butyryl group or a pivaloyl group; a lower haloalkanoyl group such asa trifluoroacetyl group; a lower alkoxycarbonyl group such as amethoxycarbonyl group, an ethoxycarbonyl group, a propoxycarbonyl groupor a tert-butoxycarbonyl group; a lower haloalkoxycarbonyl group such asa 2,2,2-trichloroethoxycarbonyl group; an alkenyloxycarbonyl group suchas a 2-propenyloxycarbonyl group; an aralkyloxycarbonyl group such as abenzyloxycarbonyl group or a p-nitrobenzyloxycarbonyl group; or a loweralkylsilyl group such as a trimethylsilyl group or atert-butyldimethylsilyl group, preferably an acetyl group, atrifluoroacetyl group, a tert-butoxycarbonyl group or abenzyloxycarbonyl group.

A protecting group for a carboxyl group is a lower alkyl group such as amethyl group, an ethyl group, a propyl group, an isopropyl group or atert-butyl group; a lower haloalkyl group such as a 2,2,2-trichloroethylgroup; a lower alkenyl group such as a 2-propenyl group; or an aralkylgroup such as a benzyl group, a p-methoxybenzyl group, a p-nitrobenzylgroup, a benzhydryl group or a trityl group, preferably, a methyl group,an ethyl group, a tert-butyl group, a 2-propenyl group, a benzyl group,a p-methoxybenzyl group or a benzhydryl group.

A salt of the compound represented by general formula (I) may be apharmaceutically acceptable common salt, which may be a base-additionsalt resulting from addition of a base to an acidic group, if any, suchas a carboxyl group at the 3a-position or any other acidic group, or anacid-addition salt resulting from addition of an acid to an amino group,if any, or a basic heterocyclic ring, if any.

The base-addition salt may, for example, be an alkali metal salt such asa sodium salt or a potassium salt; an alkaline earth metal salt such asa calcium salt or a magnesium salt; an ammonium salt; or an organicamine salt such as a trimethylamine salt, a triethylamine salt, adicyclohexylamine salt, an ethanolamine salt, a diethanolamine salt, atriethanolamine salt, a procaine salt or an N,N'-dibenzylethylenediaminesalt.

The acid-addition salt may, for example, be an inorganic acid salt suchas a hydrochloride, a sulfate, a nitrate, a phosphate or a perchlorate;an organic acid salt such as a maleate, a fumarate, a tartrate, acitrate, an ascorbate or a trifluoroacetate; or a sulfonate such as amethanesulfonate, an isethionate, a benzenesulfonate or ap-toluenesulfonate.

An ester of the compound represented by general formula (I) may be apharmaceutically acceptable common ester resulting from esterificationof a carboxyl group at the 3a-position or any other carboxyl group, andmay, for example, be an ester with a lower alkyl group such as a methylgroup, an ethyl group, a propyl group, an isopropyl group, a butylgroup, a sec-butyl group, a tert-butyl group, a pentyl group, anisopentyl group, a neopentyl group, a cyclopropyl group, a cyclobutylgroup or a cyclopentyl group, an ester with aralkyl group such as abenzyl group or a phenethyl group, an ester with a lower alkenyl groupsuch as an allyl group or a 2-butenyl group, an ester with a loweralkoxyalkyl group such as a methoxymethyl group, a 2-methoxyethyl groupor a 2-ethoxyethyl group, an ester with a lower alkanoyloxyalkyl groupsuch as an acetoxymethyl group, a pivaloyloxymethyl group or a1-pivaloyloxyethyl group, an ester with a lower alkoxycarbonylalkylgroup such as a methoxycarbonylmethyl group or anisopropoxycarbonylmethyl group, an ester with a lower carboxyalkyl groupsuch as a carboxymethyl group, an ester with a loweralkoxycarbonyloxyalkyl group such as a 1-(ethoxycarbonyloxy)ethyl groupor a 1-(cyclohexyloxycarbonyloxy)ethyl group, an ester with a lowercarbamoyloxyalkyl group such as a carbamoyloxymethyl group, an esterwith a phthalidyl group or an ester with a(5-substituted-2-oxo-1,3-dioxol-4-yl)methyl group such as a(5-methyl-2-oxo-1,3-dioxol-4-yl)methyl group.

An azole type antifungal agent is a compound which has an antifungalactivity and an imidazole or triazole ring in its molecule, and is oneor at least two selected from antifungal agents, for example, disclosedin Clinical Infectious Diseases, vol. 14 (Suppl 1), S161-9 (1992) suchas butoconazole, oxiconazole, clotrimazole, terconazole, econazole,tioconazole, miconazole, fluconazole, ketoconazole and itraconazole,preferably miconazole, fluconazole or itraconazole. These agents arecommercially available or obtainable in accordance with theabove-mentioned reference.

The compounds represented by general formula (I) include the compoundsrepresented by general formula (I-a): ##STR3## wherein R^(1a) is ahydrogen atom, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which is not substituted, a C₁ -C₁₆ alkyl group, a C₂-C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇-C₁₅ aralkyl group or a heterocyclic group which has one to fivesubstituents selected from the group consisting of halogen atoms, cyanogroups, hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆alkylcarbonyloxy groups, amino groups, mono-C₁ -C₁₆ alkylamino groups,di-C₁ -C₁₆ alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonylgroups, aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂aryloxy groups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or agroup represented by --Y^(a) --R^(3a) ; Y^(a) is a carbonyl group, athiocarbonyl group or a sulfonyl group; R^(3a) is a C₁ -C₁₆ alkyl group,a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup, a C₇ -C₁₅ aralkylamino group or a heterocyclic group which is notsubstituted, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or aheterocyclic group which has one to four substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, aminogroups and carboxyl groups, and hydroxyl groups, amino groups andcarboxyl groups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkylgroup, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, acarboxy-C₁ -C₁₆ alkyl group or a protecting group; R_(2a) is a C₁ -C₁₆alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆ -C₁₂aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic group which is notsubstituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which has one to five substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, C₁-C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxy groups, amino groups,mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆ alkylamino groups, carboxylgroups, C₁ -C₁₆ alkyloxycarbonyl groups, aminocarbonyl groups, sulfogroups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxy groups, C₇ -C₁₅ aralkyloxygroups and heterocyclic groups, or a group represented by --Y^(b)--R^(3b) ; Y^(b) is a carbonyl group, a thiocarbonyl group or a sulfonylgroup; and R^(3b) is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, aC₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆-C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino groupor a heterocyclic group which is not substituted, or a C₁ -C₁₆ alkylgroup, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup, a C₇ -C₁₅ aralkylamino group or a heterocyclic group which hasone to four substituents selected from the group consisting of halogenatoms, cyano groups, hydroxyl groups, amino groups and carboxyl groups,and hydroxyl groups, amino groups and carboxyl groups having a C₁ -C₁₆alkyl group, a halo-C₁ -C₁₆ alkyl group, a hydroxy-C₁ -C₁₆ alkyl group,an amino-C₁ -C₁₆ alkyl group, a carboxy-C₁ -C₁₆ alkyl group or aprotecting group (provided that when Y^(b) is a carbonyl group, andR^(3b) is a C₁ -C₁₆ alkyl group which is not substituted, R^(1a) is nota C₁ -C₁₆ alkyl group which is not substituted), the compoundsrepresented by general formula (I-b): ##STR4## wherein R^(1b) is a C₁-C₁₆ alkyl group which is not substituted; R^(2b) is a group representedby --Y^(c) --R^(3c) ; Y^(c) is a carbonyl group; and R^(3c) is a C₁ -C₁₆alkyl group which is not substituted (provided that both R^(1b) andR^(3c) are not methyl groups at the same time), the compoundsrepresented by general formula (I-c): ##STR5## wherein R^(1c) is ahydrogen atom, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆-C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic group, or aC₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, aC₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic groupwhich has one to five substituents selected from the group consisting ofhalogen atoms, cyano groups, hydroxyl groups, C₁ -C₁₆ alkyloxy groups,C₁ -C₁₆ alkylcarbonyloxy groups, amino groups, mono-C₁ -C₁₆ alkylaminogroups, di-C₁ -C₁₆ alkylamino groups, carboxyl groups, C₁ -C₁₆alkyloxycarbonyl groups, aminocarbonyl groups, sulfo groups, C₆ -C₁₂aryl groups, C₆ -C₁₂ aryloxy groups, C₇ -C₁₅ aralkyloxy groups andheterocyclic groups, or a group represented by --Y--R³ ; Y is a carbonylgroup, a thiocarbonyl group or a sulfonyl group; R³ is a C₁ -C₁₆ alkylgroup, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup, a C₇ -C₁₅ aralkylamino group or a heterocyclic group which is notsubstituted, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or aheterocyclic group which has one to four substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, aminogroups and carboxyl groups, and hydroxyl groups, amino groups andcarboxyl groups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkylgroup, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, acarboxy-C₁ -C₁₆ alkyl group or a protecting group; and R^(2c) is ahydrogen atom (provided that when Y is a carbonyl group, R³ is not a C₁-C₁₆ alkyl group which is not substituted), the compounds represented bygeneral formula (I-d): ##STR6## wherein R^(1d) is a C₁ -C₁₆ alkyl groupwhich is not substituted; and R^(2c) is a hydrogen atom (provided thatR^(1d) is not a methyl group) and the compounds represented by generalformula (I-e): ##STR7## wherein R^(1e) is a group represented by --Y^(c)--R^(3d) ; Y^(c) is a carbonyl group; R^(3d) is a C₁ -C₁₆ alkyl groupwhich is not substituted; and R^(2c) is a hydrogen atom (provided thatR^(3d) is not a methyl group).

Among the compounds represented by general formula (I), preferred arethose wherein R¹ is a hydrogen atom, an unsubstituted C₁ -C₁₆ alkylgroup such as an ethyl group, a propyl group, an isopropyl group, abutyl group, a pentyl group, an isopentyl group, a hexyl group, a decylgroup or a cetyl group, preferably a butyl group, a pentyl group, anisopentyl group, a hexyl group or a decyl group, an unsubstituted C₂-C₁₀ alkenyl group such as a 3-methyl-2-butenyl group, an unsubstitutedC₇ -C₁₅ aralkyl group such as a benzyl group; a C₁ -C₁₆ alkyl group or aC₃ -C₆ alkynyl group which has one to five substituents selected fromthe group consisting of cyano groups, C₁ -C₁₆ alkyloxycarbonyl groups,aminocarbonyl groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxy groups, C₇-C₁₅ aralkyloxy groups and heterocyclic groups such as a 3-cyanopropylgroup, an ethoxycarbonylmethyl group, an aminocarbonylmethyl group, a3-phenoxypropyl group, a 3-benzyloxypropyl group, a 4-pyridylmethylgroup, a 2-(N-piperidino)ethyl group, a 3-(N-1H-pyrrolo)propyl group ora phenylacetyl group; or a group represented by --Y--R³ wherein Y is acarbonyl group, and R³ is an unsubstituted C₁ -C₁₆ alkyl group, anunsubstituted C₂ -C₁₀ alkenyl group, an unsubstituted C₆ -C₁₂ arylgroup, an unsubstituted C₇ -C₁₅ aralkyl group, an unsubstituted C₇ -C₁₅aralkylamino group or a C₁ -C₁₆ alkyl group having one to four carboxylgroups such as a propionyl group, a butyryl group, a valeryl group, a3-methylbutyryl group, a hexanoyl group, a heptanoyl group, a decanoylgroup, a lauroyl group or a palmitoyl group, preferably, a butyrylgroup, a valeryl group, a hexanoyl group, a heptanoyl group or adecanoyl group, or a crotonoyl group, a 2,4-hexadienoyl group, a benzoylgroup, a 3-phenylpropionyl group, a benzylaminocarbonyl group or a3-carboxypropionyl group, R² is a hydrogen atom or a group representedby --Y--R³ wherein Y is a carbonyl group, and R³ is an unsubstituted C₁-C₁₆ alkyl group, an unsubstituted C₆ -C₁₂ aryl group or a C₁ -C₁₆ alkylgroup having one to four carboxyl groups such as a propionyl group, avaleryl group, a heptanoyl group, a decanoyl group, a benzoyl group or a3-carboxypropionyl group.

Among the compounds represented by general formula (I-d), preferred arethose wherein R^(1d) is an unsubstituted C₄ -C₁₀ alkyl group,particularly a butyl group, a pentyl group, an isopentyl group, a hexylgroup or a decyl group.

Among the compounds represented by general formula (I-e), preferred arethose wherein R^(3d) is an unsubstituted C₃ -C₉ alkyl group,particularly a propyl group, a butyl group, a pentyl group, a hexylgroup or a nonyl group.

The compounds of the present invention represented by general formula(I) can be prepared, for example, by the following process.

Namely, the compounds of the present invention can be prepared bychemical modification of BE-31405 as the starting material into acompound represented by general formula (I) wherein R¹ or R² is ahydrogen atom and then introducing a substituent corresponding to R¹ orR² in a compound of the present invention instead of the hydrogen atom.

In the preparation, it is preferred to protect a functional group whichdoes not participate in the reaction, if necessary, and deprotect itafter the reaction.

For introduction of these substituents, well-known chemical techniquessuch as alkylation, alkenylation, aralkylation, alkanoylation,arylation, thiocarbonylation and sulfonylation may be used.

These terms should be interpreted broadly and cover any reactions forintroduction of substituents corresponding to R¹ and R² in a compoundrepresented by general formula (I) of the present invention. Forexample, alkanoylation means introduction of a substituted orunsubstituted alkanoyl group defined in the present invention.

Alkylation, alkenylation, alkynylation or aralkylation of a compoundwherein R¹ or R² is a hydrogen atom can be accomplished in accordancewith known methods using, for example, an alkylation, alkenylation,alkynylation or aralkylation agent such as an alkyl halide, an alkenylhalide, an alkynyl halide, an aralkyl halide, an alkyl mesylate, analkenyl mesylate, an aralkyl mesylate, an alkyl tosylate or an aralkyltosylate.

Alkylation, alkenylation, alkynylation or aralkylation of a compoundwherein R¹ or R² is a hydrogen atom can be accomplished by treating thecompound wherein R¹ or R² is a hydrogen atom with an alkylation,alkenylation, alkynylation or aralkylation agent in an appropriatesolvent.

The solvent may be dimethylformamide, methylene chloride, dimethylsulfoxide or a mixture thereof.

The reaction temperature is usually within a range of from about -20° C.to the boiling point of the solvent, preferably from 20° C. to 60° C.,though it may be below the range, if necessary.

The reaction time is usually from 10 minutes to 24 hours, preferablyfrom 1 hour to 12 hours, though it may be longer or shorter, ifnecessary.

The amount of an alkylation, alkenylation, alkynylation or aralkylationagent used for a compound wherein R¹ or R² is a hydrogen atom is usuallyat least 1 mole, preferably from 1 to 10 moles, more preferably from 2to 5 moles in relation to the compound wherein R¹ or R² is a hydrogenatom, though it may be varied widely according to the kind of thecompound or the reaction conditions without any restriction.

Alkanoylation or alkylthiocarbonylation of a compound wherein R¹ or R²is a hydrogen atom can be accomplished by treating the compound whereinR¹ or R² is a hydrogen atom with an acid halide or an acid anhydridecorresponding to a given substituent in an appropriate solvent.

The solvent may be dimethylformamide, pyridine, methylene chloride,dimethyl sulfoxide or a mixture thereof.

The reaction temperature is usually within a range of from about -5° C.to the boiling point of the solvent, preferably from 20° C. to 60° C.,though it may be below the range, if necessary.

The reaction time is usually from 30 minutes to 2 days, preferably from1 hour to 24 hours, though it may be longer or shorter, if necessary.

The amount of an acid halide or an acid anhydride used for a compoundwherein R¹ or R² is a hydrogen atom is usually at least 1 mole,preferably from 1 to 5 moles, more preferably from 1 to 3 moles inrelation to the compound wherein R¹ or R² is a hydrogen atom, though itmay be varied widely according to the kind of the compound or thereaction conditions without any restriction.

Sulfonylation of a compound wherein R¹ or R² is a hydrogen atom can beaccomplished by treating the compound wherein R¹ or R² is a hydrogenatom with an organic sulfonyl halide or an organic sulfonic anhydridecorresponding to a given substituent in an appropriate solvent in thepresence or absence of a base.

The solvent may be dimethylformamide, methylene chloride, dimethylsulfoxide or a mixture thereof.

The base may be sodium hydride or lithium hydride.

The reaction temperature is usually within a range of from about -10° C.to about 50° C., preferably from 20° C. to 60° C., though it may bebelow the range, if necessary.

The reaction time is usually from 30 minutes to 3 days, preferably from1 hour to 24 hours, though it may be longer or shorter, if necessary.

The amount of an organic sulfonyl halide or an organic sulfonicanhydride used for a compound wherein R¹ or R² is a hydrogen atom isusually a small excess, preferably from 1 to 3 moles in relation to thecompound wherein R¹ or R² is a hydrogen atom, though it may be variedwidely according to the kind of the compound or the reaction conditionswithout any restriction.

In the above-mentioned methods, the protecting groups for functionalgroups which do not participate in the reaction may be protecting groupsfor a hydroxyl group, protecting groups for an amino group andprotecting groups for a carboxyl group as described above or the like.

Introduction and elimination of the protecting groups can beaccomplished by the methods disclosed in the literature (ProtectiveGroups in Organic Synthesis, written by T. W. Greene, published by JohnWiley & Sons (1981)) or similar methods or any ordinary methods widelyknown in the field of chemistry.

The compounds produced by the above-mentioned reactions can be isolatedor purified through techniques already known in the field of organicchemistry such as precipitation, extraction with solvent,recrystallization and chromatography.

The compounds produced by the above-mentioned reactions can be convertedinto pharmaceutically acceptable salts or esters thereof or vice versaby ordinary methods.

The starting material, BE-31405, can be obtained, for example, by usingmicroorganisms such as Penicillium sp. F-31405, as disclosed inJP-A-6-157582, or its mutant strain more productive of the compound,Penicillium sp. F31405-17M.

Penicillium sp. F31405-17M has the following mycologicalcharacteristics.

(1) Morphology

The F31405-17M strain is the same in shape as its origin, Penicilliumsp. F-31405, with conidiophores of 110 to 210×1.8 to 3.6 μm and a smoothsurface or fine projections and forms symmetrical double verticillatepenicilli. The metulae are 10.0 to 13.1×2.3 to 3.1 μm and grow inbunches of 4 to 8. The phialides are (9.7 to) 11.4 to 15.0×1.8 to 2.6 μmand verticillate. The conidia have smooth surfaces and are subsphericalor elliptic or ovoid and 3.5 to 4.4×2.6 to 3.5 μm in size.

(2) Culture Characteristics

The culture characteristics of the F31405-17M strain are slightlydifferent from those of its origin F-31405. Table 1 shows the growthcharacteristics observed after 7 days of incubation on various agarmedia at 25° C. The colors in the Table were identified on the basis ofthe names of colors in Methuen Handbook of Color, 3rd ed., (1984).

                  TABLE 1                                                         ______________________________________                                        Growth characteristics of the F31405-17M strain                                      Diameter                                                                      of                   Color of                                          Culture                                                                              colonies  Color of   the colony                                                                             Colony                                   medium (mm)      colonies   reverse  texture                                  ______________________________________                                        Czapek 13-15     Pale green Pale green                                                                             Slightly                                 agar             to grayish to grayish                                                                             velutinous                                                green      green                                             Czapek-                                                                              17-20     Yellowish  Bright   Slightly                                 yeast            white to   yellow to                                                                              velutinous                               extract          dark green yellowish                                         agar                        white                                             Malt   30-32     Dark green Yellowish                                                                              velutinous                               extract                     gray to                                           agar                        yellowish                                                                     white                                             ______________________________________                                    

The strain develops enough conidia on any culture medium, especially onthe malt extract agar and secretes nothing. The strain produces a brightyellow soluble pigment on the Czapek agar and the Czapek-yeast extractagar. The improved strain F31405-17M does not form premature sclerotiumtissue when incubated at 37° C., like its origin Penicillium sp. F-31405can.

The growth is poor at 37° C. than at 25° C. on any culture medium. Thestrain is viable in a temperature range of from 12 to 37° C. with theoptimum growth temperature of 28.5° C., and in a pH range of from 2 to11 with the optimum growth pH of about 6.5.

Penicillium sp. F-31405 and F31405-17M have been placed on internationaldeposit in National Institute of Bioscience and Human-Technology (NIBH),Agency of Industrial Science and Technology, Ministry of InternationalTrade and Industry (address: 1-3, Higashi 1-chome, Tsukuba-Shi,Ibaraki-ken 305, Japan) with the deposition numbers of FERM BP-5714(date of original deposition: Oct. 20, 1992) and FERM BP-5716 (date oforiginal deposition: Sep. 13, 1996), respectively.

Compounds of the present invention represented by general formula (I)have excellent antifungal actions, as is evident from the biologicalactivities shown below. The actifungal effects against a fungus areshown in Table 2.

                  TABLE 2                                                         ______________________________________                                        Biological activities (antifungal activities) of BE-31405 derivatives         Compound No..sup.a                                                                           MIC (μg/ml).sup.b                                           ______________________________________                                        Compound 10    6.25                                                           Compound 13    3.13                                                           Compound 14    1.56                                                           Compound 38    3.13                                                           Compound 41    1.56                                                           Compound 44    3.13                                                           Compound 47    1.56                                                           Compound 50    3.13                                                           ______________________________________                                         .sup.a Under the same experimental conditions, the MIC of BE31405 in          single use was 50 μg/ml, and the MIC of SCH57404 (xylarin) in single       use was 100 μg/ml.                                                         .sup.b MICs were determined against Candida albicans IFO1385 after 2 days     of incubation of the test strain at 37° C. on yeastnitrogen base       agar (containing 1% glucose and 0.25% dipotassium hydrogen phosphate,         Difco).                                                                  

Although as shown in Table 2, BE-31405 shows growth inhibitory activityagainst a certain kind of fungus in vitro especially under acidicconditions, the compound is severely restricted in practical medical useby its big drawback that the activity remarkably lowers under neutralconditions, considering that the pH of blood is around neutrality.

The compounds of the present invention are excellent compounds showingstrong antifungal activities under neutral conditions imparted bychemical modification to BE-31405 as well as under acidic conditions andare quite useful as antifungal agents.

Combined use of an azole type antifungal agent with the compounds of thepresent invention additively and synergically intensifies the antifungalactivities of the compounds of the present invention.

In the case of the above-mentioned combined use, a compound of thepresent invention and an azole type antifungal agent may be administeredsimultaneously or separately or after formulation of a compositioncontaining them as a pharmaceutical.

The compound or antifungal composition of the present invention can beadministered orally or parenterally in its clinical application, and itmay be formulated to meet the administration mode by adding variouspharmaceutically acceptable additives, as the case requires, and used asan antifungal agent.

The form for such formulation may, for example, be solid formulationssuch as tablets, capsules, granules, pills, troches, powders orsuppositories, or liquid formulations such as syrups, elixirs,suspensions or injections, as well as aerosols, eyedrops, ointments,ophthalmic ointments, emulsions, creams, liniments or lotions. Theseformulations may be prepared in accordance with conventional methodscommonly used in the field of drug formulations.

As the additives, various additives which are commonly used in the drugformulation field, can be used. For example, saccharides such as lactoseor glucose, a starch from corn, wheat or rice, a vegetable oil such assoybean oil, peanuts oil or sesame oil, a fatty acid such as stearicacid, an inorganic salt such as magnesium metasilicate aluminate oranhydrous calcium phosphate, a synthetic polymer such aspolyvinylpyrrolidone or polyalkylene glycol, a fatty acid salt such ascalcium stearate or magnesium stearate, an alcohol such as stearylalcohol or benzyl alcohol, a synthetic cellulose derivative such asmethyl cellulose, carboxymethyl cellulose, ethyl cellulose orhydroxy-propylmethyl cellulose, or others such as water, gelatin, talcand gum arabic, may, for example, be mentioned.

Further, in the case of a liquid formulation, it may be in such a formthat at the time of use, it is dissolved or suspended in water or inother suitable medium. Especially when administration is carried out bye.g. intramuscular injection, intravenous injection or subcutaneousinjection, a suitable medium for such an injection may, for example, bedistilled water for injection, a lidocaine hydrochloride aqueoussolution (for intramuscular injection), physiological saline, an aqueousglucose solution, ethanol, liquid for intravenous injection (such as anaqueous solution of citric acid and sodium citrate) or an electrolytesolution (for intravenous drip and intravenous injection), or a mixedsolution thereof. Further, a buffer or a preservative may be added.

These formulations may contain usually from 0.1 to 100 wt %, preferablyfrom 5 to 100 wt %, of the active ingredient in the case of theabove-mentioned solid formulations, and may contain from 0.1 to 10 wt %,preferably from 1 to 5 wt %, in the case of other formulations.

When the compounds of the present invention are used together with anazole type antifungal agent as a composition, the weight ratio of acompound represented by general formula (I) or a pharmaceuticallyacceptable salt or ester thereof to an azole type antifungal agent isfrom 0.001:1 to 1000:1, preferably 0.05:1 to 20:1.

A practically preferred dose of the compound or the antifungalcomposition of the present invention varies depending upon the type ofthe compound used, the type of the composition blended, the sex, age,weight, diseased degree and the particular section to be treated of thepatient, but it is usually from 0.1 to 100 mg/kg in the case of oraladministration and from 0.01 to 100 mg/kg in the case of parenteraladministration, per adult per day. The number of times of administrationvaries depending upon the administration method and the symptom, but itis preferred to carry out the administration from one to five times perday.

As described above, the present invention provides a useful antifungalagent and, needless to say, a novel treatment for mycosis.

BEST MODE FOR CARRYING OUT THE INVENTION

Now, the present invention will be described in further detail withreference to Examples, Formulation Examples and Reference Examples, butthe present invention is by no means restricted thereby.

In the following Examples, BE-31405 obtained in Reference Examples isreferred to as compound (1), and a compound wherein R¹ and R² arehydrogen is referred to as compound (2).

EXAMPLE 1

Preparation of8a-[[[6-(hydroxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (2)

1.02 g of compound (1) was dissolved in 60 ml of methanol, mixed with 28ml of 0.1 N aqueous NaOH and allowed to react at room temperature for 50minutes under stirring. The reaction solution was poured into 600 ml of0.1 M sodium phosphate buffer (pH 5.57), and the product was extractedwith 600 ml of ethyl acetate. The ethyl acetate layer was washed with400 ml and 300 ml portions of water. The washed ethyl acetate layer wasconcentrated in vacuo to give 871.8 mg of compound (2) as a colorlesspowder.

Rf: 0.37 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 513[M+Na]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.13 (1H, brd, J=3.7 Hz),5.73(1H, brd, J=3.0 Hz), 4.49(1H, m), 4.24(1H, m), 4.06(1H, t, J=3.7Hz), 4.02(1H, d, J=9.8 Hz), 3.98(1H, d, J=9.8 Hz), 3.71(1H, brs),2.82(1H, t, J=3.7 Hz), 2.34(1H, m), 1.95-2.12(5H, m), 1.86(1H, m), 1.76(2H, m), 1.31(1H, d, J=12.8 Hz), 1.23(1H, m), 1.04(3H, d, J=6.7 Hz),1.03(1H, m), 0.97(3H, d, J=6.7 Hz), 0.79 (3H, d, J=7.0 Hz).

EXAMPLE 2

Preparation of8a-[[[6-(propionyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (7)

15 mg of compound (2) was dissolved in 1.0 ml of pyridine and stirredtogether with 7.9 μl of propionic anhydride at room temperature for 92hours.

The reaction solution was concentrated in vacuo and charged onto asilica gel column (Kieselgel 60, Merck, 1.5φ×27 cm) and eluted withchloroform-methanol (50:1). The fraction containing the desired productwas concentrated in vacuo to give 5.1 mg of compound (7) as a colorlessoily substance.

Rf: 0.45 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 547[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.75(1H, brd, J=3.0 Hz), 4.75(1H, m), 4.71(1H, t, J=4.0 Hz), 4.39(1H,m), 4.01(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.69(1H, brs),2.80(1H, t, J=3.7 Hz), 2.45(2H, m), 2.33(1H, m), 1.90-2.10(5H, m),1.87(1H, m), 1.76(2H, m), 1.30(1H, d, J=12.5 Hz), 1.23(1H, m), 1.19(3H,t, J=7.6 Hz), 1.03(3H, d, J=6.7 Hz), 1.02(1H, m), 0.96(3H, d, J=6.7 Hz),0.79(3H, d, J=6.7 Hz).

EXAMPLE 3

Preparation of8a-[[[6,7-di(propionyloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (8)

In the silica gel chromatography in Example 2, the fraction containingthe desired compound eluted earlier than compound (7) was concentratedto give 2.0 mg of compound (8).

Rf: 0.67 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 603[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.63(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.83(1H, brd, J=3.0 Hz), 5.02(1H, brs), 4.78(1H, m), 4.73(1H, t, J=4.0Hz), 4.44(1H, m), 4.04(1H, d, J=9.8 Hz), 3.92(1H, d, J=9.8 Hz), 2.67(1H,t, J=3.7 Hz), 2.47(4H, m), 2.32(1H, m), 1.83-2.10(6H, m), 1.73(2H, m),1.28(1H, d, J=12.8 Hz), 1.23(1H, m), 1.20(6H, t, J=7.6 Hz), 1.02(3H, d,J=6.7 Hz), 1.01(1H, m), 0.95(3H, d, J=6.7 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 4

Preparation of8a-[[[6-(n-butyryloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (9)

300 mg of compound (2) was dissolved in 1.5 ml of dry pyridine andstirred together with 130 μl of n-butyric anhydride at room temperaturefor 5 hours. The reaction solution was concentrated in vacuo, chargedonto a silica gel column (Kieselgel 60, Merck, 1.5φ×30 cm) and elutedwith 700 ml of n-hexane-ethyl acetate (1:1) and then with ethyl acetate.The fraction containing the desired product was concentrated in vacuo togive the crude desired compound. For further purification, it wassubjected to silica gel column chromatography (Kieselgel 60, Merck,1.5φ×30 cm) and eluted with chloroform-methanol (50:1). The fractioncontaining the desired product was concentrated to dryness to give 106mg of compound (9).

Rf: 0.62 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 561[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.09 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=3.0 Hz), 4.76(1H, m), 4.70(1H, dd, J=4.6, 3.3 Hz),4.39(1H, m), 3.99(1H, d, J=9.8 Hz), 3.97(1H, d, J=9.8 Hz), 3.68(1H, d,J=1.5 Hz), 2.79(1H, t, J=3.7 Hz), 2.40(2H, m), 2.32(1H, m),1.90-2.10(5H, m), 1.87(1H, m), 1.75(2H, m), 1.70(2H, m), 1.29(1H, d,J=12.5 Hz), 1.23(1H, m), 1.02(3H, d, J=6.7 Hz), 1.02(1H, m), 0.99(3H, t,J=7.3 Hz), 0.95(3H, d, J=6.7 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 5

Preparation of8a-[[[6-(n-valeryloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (10)

20 mg of compound (2) was dissolved in 1.0 ml of pyridine and stirredtogether with 30.4 μl of n-valeric anhydride at room temperature. After55 hours, 16.1 μl of n-valeric anhydride was added, and the reaction wasconducted for another 17 hours. The reaction solution was concentratedin vacuo, charged onto a silica gel column (Kieselgel 60, Merck, 1.5φ×30cm) and eluted with chloroform-methanol (100:1). The fraction containingthe desired product was concentrated in vacuo to give the desiredcompound as a crude product.

For further purification, it was subjected to Sephadex LH-20 columnchromatography (Pharmacia, 1.5φ×30 cm) and eluted with methanol. Thefraction containing the desired product was concentrated to dryness togive 10.3 mg of compound (10).

Rf: 0.39 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 575[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.09 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.76(1H, m), 4.69(1H, t, J=3.7 Hz), 4.39(1H,m), 4.00(1H, d, J=9.8 Hz), 3.97(1H, d, J=9.8 Hz), 3.68(1H, brs),2.80(1H, t, J=3.7 Hz), 2.42(2H, m), 2.33(1H, m), 1.92-2.12(5H, m),1.85(1H, m), 1.76(2H, m), 1.64(2H, m), 1.38(2H, m), 1.30(1H, d, J=12.5Hz), 1.24(1H, m), 1.02(3H, d, J=6.7 Hz), 1.01(1H, m), 0.96(3H, d, J=6.7Hz), 0.94(3H, t, J=7.3 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 6

Preparation of8a-[[[6,7-di(n-valeryloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (11)

In the silica gel chromatography in Example 5, the fraction containingthe desired compound eluted earlier than compound (10) was concentratedto give 8.0 mg of compound (11).

Rf: 0.52 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 659[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.83(1H, brd, J=3.0 Hz), 5.01(1H, brs), 4.78(1H, m), 4.71(1H, t, J=4.0Hz), 4.42(1H, m), 3.98(1H, d, J=9.8 Hz), 3.94(1H, d, J=9.8 Hz), 2.67(1H,t, J=3.7 Hz), 2.45(4H, m), 2.32(1H, m), 1.82-2.10(6H, m), 1.60-1.78(6H,m), 1.40(4H, m), 1.28(1H, d, J=12.8 Hz), 1.22(1H, m), 1.02(3H, d, J=6.7Hz), 1.01(1H, m), 0.96(6H, d, J=7.6 Hz), 0.95(3H, d, J=6.7 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 7

Preparation of8a-[[[6-(3-methylbutyryloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (12)

41.4 mg of compound (2) was dissolved in 0.3 ml of dry pyridine andstirred together with 14.2 μl of isovaleric anhydride at roomtemperature. After 43 hours, 7.1 μl of isovaleric anhydride was added,and the reaction was conducted for another 26 hours. The reactionsolution was concentrated in vacuo, mixed with 2 ml of methanol and 0.2ml of water and washed with 2 ml of n-hexane twice and with petroleumether twice. The methanol layer was concentrated in vacuo, and thereaction product was dissolved in 1 ml of chloroform, charged onto asilica gel column (Kieselgel 60, Merck, 1.5φ×25 cm) and eluted with 150ml of chloroform and then with chloroform-methanol (50:1). The fractioncontaining the desired product was concentrated in vacuo to give 10.6 mgof compound (12) as a colorless oily substance.

Rf: 0.42 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 575[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.08 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.78(1H, m), 4.68(1H, t, J=3.7 Hz), 4.39(1H,m), 3.99(1H, d, J=9.8 Hz), 3.95(1H, d, J=9.8 Hz), 3.68(1H, d, J=1.5 Hz),2.80(1H, t, J=3.7 Hz), 2.31(3H, m), 1.83-2.16(6H, m), 1.75(2H, m),1.30(1H, d, J=12.5 Hz), 1.23(1H, m), 1.02(3H, d, J=6.7 Hz), 1.02(1H, m),1.00(6H, d, J=6.7 Hz), 0.94(3H, d, J=6.7 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 8

Preparation of8a-[[[6-(n-hexanoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (13)

39.0 mg of compound (2) was dissolved in 0.3 ml of dry pyridine andstirred together with 15.6 μl of n-hexanoic anhydride at roomtemperature. After 43 hours, 7.8 μl of n-hexanoic anhydride was added,and the reaction was conducted for another 26 hours. The reactionsolution was concentrated in vacuo, mixed with 1 ml of methanol and 0.1ml of water and washed with 1.5 ml of n-hexane twice and with petroleumether twice. The methanol layer was concentrated in vacuo, charged ontoa silica gel column (Kieselgel 60, Merck, 1.5φ×25 cm) and eluted with150 ml of chloroform and then with chloroform-methanol (50:1). Thefraction containing the desired product was concentrated in vacuo togive 15.1 mg of compound (13) as a colorless oily substance.

Rf: 0.46 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 589[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.09 (1H, brd, J=3.7 Hz),5.75(1H, brd, J=4.0 Hz), 4.75(1H, m), 4.70(1H, t, J=3.7 Hz), 4.39(1H,m), 3.99(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.68(1H, d, J=1.5 Hz),2.81(1H, t, J=3.7 Hz), 2.41(2H, m), 2.32(1H, m), 1.84-2.12(6H, m),1.75(2H, m), 1.66(2H, m), 1.33(4H, m), 1.30(1H, d, J=12.5 Hz), 1.23(1H,m), 1.02(3H, d, J=6.7 Hz), 1.00(1H, m), 0.94(3H, d, J=6.7 Hz), 0.91(3H,t, J=7.3 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 9

Preparation of8a-[[[6-(n-heptanoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (14)

30 mg of compound (2) was dissolved in 1.0 ml of pyridine and stirredtogether with 54.7 μl of n-heptanoic anhydride at room temperature.After 42 hours, 29.6 μl of n-heptanoic anhydride was added, and thereaction was conducted for another 7 hours. The reaction solution wasconcentrated in vacuo, charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×30 cm) and eluted with chloroform-methanol (100:1). Thefraction containing the desired product was concentrated in vacuo togive the desired compound as a crude product. For further purification,it was subjected to Sephadex LH-20 column chromatography (Pharmacia,1.5φ×90 cm) and eluted with methanol. The fraction containing thedesired product was concentrated in vacuo to dryness to give 12.1 mg ofcompound (14).

Rf: 0.66 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 603[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃),:9.70(1H, s), 6.09 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.75(1H, m), 4.70(1H, t, J=3.7 Hz), 4.39(1H,m), 3.99(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.68(1H, d, J=1.5 Hz),2.80(1H, t, J=3.7 Hz), 2.41(2H, m), 2.32(1H, m), 1.82-2.10(6H, m),1.74(2H, m), 1.64(2H, m), 1.20-1.40 (8H, m), 1.02(3H, d, J=6.7 Hz),1.00(1H, m), 0.95(3H, d, J=6.7 Hz), 0.90(3H, t, J=7.3 Hz), 0.79(3H, d,J=6.7 Hz).

EXAMPLE 10

Preparation of8a-[[[6,7-di(n-heptanoyloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (15)

In the silica gel chromatography in Example 9, the fraction containingthe desired compound eluted earlier than compound (14) was concentratedto give the desired compound as a crude product.

For further purification, it was subjected to Sephadex LH-20 columnchromatography (Pharmacia, 1.5φ×30 cm) and eluted with methanol. Thefraction containing the desired product was concentrated in vacuo todryness to give 8.3 mg of compound (15).

Rf: 0.80 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 715[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃),:9.63(1H, s), 6.08 (1H, brd, J=3.7 Hz),5.82(1H, brd, J=3.0 Hz), 5.00(1H, brs), 4.78(1H, m), 4.71(1H, t, J=4.0Hz), 4.41(1H, m), 3.98(1H, d, J=9.8 Hz), 3.92(1H, d, J=9.8 Hz), 2.67(1H,t, J=3.7 Hz), 2.43(4H, m), 2.32(1H, m), 1.82-2.10(6H, m), 1.58-1.78(6H,m), 1.20-1.40(14H, m), 1.02(3H, d, J=6.7 Hz), 1.00(1H, m), 0.92(3H, d,J=6.7 Hz), 0.88(6H, t, J=7.3 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 11

Preparation of8a-[[[6-(decanoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (16)

40 mg of compound (2) was dissolved in 0.5 ml of dry pyridine andstirred together with 40 mg of decanoic anhydride at room temperaturefor 72 hours. The reaction solution was concentrated in vacuo, chargedonto a silica gel column (Kieselgel 60, Merck, 1.5φ×30 cm) and elutedwith 500 ml of n-hexane-ethyl acetate (2:1) and 300 ml ofchloroform-methanol (50:1) successively. The fraction containing thedesired product was concentrated in vacuo to give the desired compoundas a crude product.

For further purification, it was subjected to silica gel columnchromatography (Kieselgel 60, Merck, 1.5φ×30 cm) and eluted withchloroform-methanol (50:1). The fraction containing the desired productwas concentrated in vacuo to dryness to give 7.9 mg of compound (16).

Rf: 0.42 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 645[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.75(1H, m), 4.70(1H, t, J=3.7 Hz), 4.39(1H,m), 4.03(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.68(1H, brs),2.80(1H, t, J=3.7 Hz), 2.41(2H, m), 2.33(1H, m), 1.94-2.12(5H, m),1.86(1H, m), 1.77(2H, m), 1.66(2H, m), 1.20-1.40(14H, m), 1.03(3H, d,J=6.7 Hz), 1.03(1H, m), 0.96(3H, d, J=6.7 Hz), 0.88(3H, t, J=7.3 Hz),0.79(3H, d, J=6.7 Hz).

EXAMPLE 12

Preparation of8a-[[[6,7-di(decanoyloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (17)

In the first silica gel chromatography in Example 11, the fractioncontaining the desired compound eluted earlier than compound (16) wasconcentrated to give 5.1 mg of compound (17).

Rf: 0.57 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 799[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.63(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.82(1H, brd, J=3.0 Hz), 5.00(1H, brs), 4.78(1H, m), 4.71(1H, t, J=4.0Hz), 4.42(1H, m), 4.02(1H, d, J=9.8 Hz), 3.92(1H, d, J=9.8Hz), 2.67(1H,t, J=3.7 Hz), 2.43(4H, m), 2.32(1H, m), 1.82-2.10(6H, m), 1.60-1.80(6H,m), 1.20-1.40(26H, m), 1.02(3H, d, J=6.7 Hz), 1.00(1H, m), 0.94(3H, d,J=6.7 Hz), 0.88(6H, t, J=7.3 Hz), 0.78(3H, d, J=6.7 Hz).

EXAMPLE 13

Preparation of8a-[[[6-(lauroyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (18)

49.8 mg of compound (2) was dissolved in 0.5 ml of dry pyridine andstirred together with 58.3 mg of lauric anhydride at room temperaturefor 24 hours. The reaction solution was concentrated in vacuo, chargedonto a silica gel column (Kieselgel 60, Merck, 1.5φ×30 cm) and elutedwith 800 ml of n-hexane-ethyl acetate-chloroform (2:1:1), 300 ml ofn-hexane-ethyl acetate-chloroform (1:1:1) and 300 ml ofchloroform-methanol (20:1) successively. The fraction containing thedesired product was concentrated in vacuo to give the desired compoundas a crude product.

For further purification, it was subjected to chromatography using asilica gel column (Kieselgel 60, Merck, 1.5φ×30 cm) and eluted withchloroform-methanol (50:1). The fraction containing the desired productwas concentrated in vacuo to dryness to give 24 mg of compound (18).

Rf: 0.43 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 673[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.75(1H, m), 4.70(1H, t, J=4.0 Hz), 4.39(1H,m), 4. 02(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.68(1H, d, J=1.5Hz), 2.80(1H, t, J=3.7 Hz), 2.41(2H, m), 2.33(1H, m), 1.94-2.12(5H, m),1.86(1H, m), 1.76(2H, m), 1.65(2H, m), 1.20-1.40(18H, m), 1.03(3H, d,J=6.7 Hz), 1.02(1H, m), 0.96(3H, d, J=6.7 Hz), 0.88(3H, t, J=7.3 Hz),0.79(3H, d, J=6.7 Hz).

EXAMPLE 14

Preparation of8a-[[[6-(palmitoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (19)

44.3 mg of compound (2) was dissolved in a mixture of 0.5 ml of drypyridine and 0.2 ml of dry dichloromethane and stirred together with37.8 mg of palmitic anhydride at room temperature. After 120 hours, 37.8mg of palmitic anhydride in 0.3 ml of dichloromethane was added, and thereaction was conducted for another 24 hours.

The reaction solution was concentrated in vacuo, charged onto a silicagel column (Kieselgel 60, Merck, 1.5φ×25 cm) and eluted with 440 ml ofn-hexane-ethyl acetate (2:1). The fraction containing the desiredproduct was concentrated in vacuo to give 19.3 mg of the crude desiredproduct. The crude desired product was further subjected to reversedphase chromatography (Capcell Pak C18 UG-120, Shiseido, 2.0φ×25 cm,0.03% trifluoroacetic acid-acetonitrile [1:9], 3.0 ml/min), and thefraction containing the desired product was concentrated in vacuo togive 6.2 mg of compound (19) as a colorless oily substance.

Rf: 0.46 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 729[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.74(1H, brd, J=4.0 Hz), 4.75(1H, m), 4.70(1H, t, J=4.0 Hz), 4.39(1H,m), 4.01(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.68(1H, d, J=1.5 Hz),2.80(1H, t, J=3.7 Hz), 2.41(2H, m), 2.33(1H, m), 1.92-2.12(5H, m),1.87(1H, m), 1.76(2H, m), 1.65(2H, m), 1.20-1.38(26H, m), 1.03(3H, d,J=6.7 Hz), 1.02(1H, m), 0.95(3H, d, J=6.7 Hz), 0.88(3H, t, J=7.3 Hz),0.79(3H, d, J=6.7 Hz).

EXAMPLE 15

Preparation of8a-[[[6-(3-carboxypropionyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (20)

20 mg of compound (2) was dissolved in 1.0 ml of pyridine and stirredtogether with 16.4 mg of succinic anhydride at room temperature. After52 hours, 8.2 mg of succinic anhydride was added, and the reaction wasconducted for another 20 hours. The reaction solution was concentratedin vacuo and subjected to reversed phase chromatography (Capcell Pak C18UG-120, Shiseido, 2.0φ×25 cm, 0.05% trifluoroacetic acid-acetonitrile[6:4], flow rate 10.0 ml/min, detection 210 nm) for purification, andthe desired fraction was recovered. The recovered fraction was pouredinto 100 ml of 0.1 M sodium phosphate buffer pH 5.5 and extracted with100 ml of ethyl acetate. The ethyl acetate extract was concentrated invacuo to give 4.0 mg of compound (20).

Rf: 0.26 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 591[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.13 (1H, brd, J=3.7 Hz),5.75(1H, brd, J=4.0 Hz), 4.74(2H, m), 4.41(1H, m), 3.99(1H, d, J=9.8Hz), 3.96(1H, d, J=9.8 Hz), 3.70(1H, d, J=1.5 Hz), 2.81(1H, t, J=3.7Hz), 2.74(4H, m), 2.32(1H, m), 1.94-2.12(5H, m), 1.85(1H, m), 1.76(2H,m), 1.31(1H, d, J=12.5 Hz), 1.24(1H, m), 1.03(3H, d, J=6.7 Hz), 1.02(1H,m), 0.96(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 16

Preparation of8a-[[[6,7-di(3-carboxypropionyloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (21)

In the reversed phase chromatography in Example 15, the fractioncontaining the desired compound eluted later than compound (20) waspoured into 100 ml of 0.1 M sodium phosphate buffer pH 5.5 and extractedwith 100 ml of ethyl acetate. The ethyl acetate extract was concentratedin vacuo to give 12 mg of compound (21).

Rf: 0.18 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 691[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.82(1H, brd, J=3.0 Hz), 4.98(1H, brs), 4.78(1H, t, J=4.0 Hz), 4.75(1H,m), 4.46(1H, m), 4.02(1H, d, J=9.8 Hz), 3.92(1H, d, J=9.8 Hz),2.64-2.80(9H, m), 2.32(1H, m), 1.70-2.10(8H, m), 1.20-1.40(2H, m),1.03(3H, d, J=6.7 Hz), 1.00(1H, m), 0.98 (3H, d, J=6.7 Hz), 0.78(3H, d,J=6.7 Hz).

EXAMPLE 17

Preparation of8a-[[[6-(crotonoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (22)

25 mg of compound (2) was dissolved in 1.0 ml of pyridine and stirredtogether with 31.5 mg of crotonic anhydride at room temperature for 18hours. The reaction solution was concentrated in vacuo and subjected toreversed phase chromatography (Capcell Pak C18 UG-120, Shiseido, 2.0φ×25cm, 0.05% trifluoroacetic acid-acetonitrile [5:5], flow rate 10.0ml/min, detection 210 nm) for purification, and the desired fraction wasrecovered. The recovered fraction was poured into 100 ml of 0.1 M sodiumphosphate buffer pH 5.5 and extracted with 100 ml of ethyl acetate. Theethyl acetate extract was concentrated in vacuo to give 4.0 mg ofcompound (22).

Rf: 0.46 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 559[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 7.08 (1H, dq, J=15.5, 7.0Hz), 6.11(1H, brd, J=3.7 Hz), 5.94 (1H, brd, J=15.5 Hz), 5.76(1H, brd,J=4.0 Hz), 4.77(2H, m), 4.42(1H, m), 4.02(1H, d, J=9.8 Hz), 3.95(1H, d,J=9.8 Hz), 3.70(1H, d, J=1.5 Hz), 2.81(1H, t, J=3.7 Hz), 2.32(1H, m),1.90-2.12(5H, m), 1.93(3H, brd, J=7.0 Hz), 1.87(1H, m), 1.75(2H, m),1.30(1H, d, J=12.5 Hz), 1.23(1H, m), 1.03(3H, d, J=6.7 Hz), 1.02(1H, m),0.95(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 18

Preparation of8a-[[[6-(benzoyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (23)

(Step 1) Preparation of8a-[[[6-(acetyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (3)

1.00 g of compound (1) was dissolved in 40 ml of methanol and mixed with760 mg of diphenyldiazomethane, and the reaction was conducted at roomtemperature for 13 hours under stirring. The reaction solution wasconcentrated in vacuo, charged onto a silica gel column (Kieselgel 60,Merck, 2.9φ×30 cm) and eluted with chloroform and then withchloroform-methanol (50:1). The fraction containing the desired productwas concentrated in vacuo to give 1.205 g of compound (3) as a colorlesspowder.

Rf: 0.75 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 699[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.73(1H, s), 7.26-7.44(10H, m), 6.96(1H,s), 6.09(1H, brd, J=3.7 Hz), 5.72(1H, brd, J=3.0 Hz), 4.71(2H, m),4.39(1H, m), 4.13(1H, d, J=9.8 Hz), 3.99(1H, d, J=9.8 Hz), 3.69(1H, brd,J=9.8 Hz), 2.82(1H, t, J=3.7 Hz), 2.46(1H, d, J=9.8 Hz), 2.25(1H, m),2.16(3H, s), 1.82-1.96(5H, m), 1.58(3H, m), 1.25(1H, d, J=12.8 Hz),0.98(3H, d, J=7.0 Hz), 0.93(2H, m), 0.73(3H, d, J=6.7 Hz), 0.26(3H, d,J=6.7 Hz).

(Step 2) Preparation of8a-[[[6-(hydroxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (4)

650 mg of compound (3) was dissolved in 46 ml of methanol and stirredtogether with 13.9 ml of 0.1 N aqueous NaOH at room temperature for 2hours. The reaction solution mixed with 24 ml of water and stirred at 4°C. for 2 hours. The reaction solution was filtered, and the recoveredprecipitate was washed with water and dried to give 464 mg of compound(4) as a colorless powder.

Rf: 0.28 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 657[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.73(1H, s), 7.26-7.44(10H, m), 6.97(1H,s), 6.10(1H, brd, J=3.7 Hz), 5.70(1H, brd, J=3.0 Hz), 4.56(1H, m),4.24(1H, m), 4.11(1H, d, J=9.8 Hz), 4.05(1H, m), 4.02(1H, d, J=9.8 Hz),3.70(1H, brd, J=9.8 Hz), 2.81(1H, t, J=3.7 Hz), 2.46(1H, d, J=9.8 Hz),2.44(1H, d, J=10.1 Hz), 2.26(1H, m), 1.83-1.96(5H, m), 1.58(3H, m),1.25(1H, d, J=12.8 Hz), 1.00(3H, d, J=6.7 Hz), 0.93(2H, m), 0.74(3H, d,J=6.7 Hz), 0.27(3H, d, J=6.7 Hz).

(Step 3) Preparation of Compound (23)

40 mg of compound (4) was dissolved in 4 ml of dichloromethane andstirred together with 1.9 mg of sodium hydride at room temperature.After 0.5 hour 13.9 mg of benzoyl chloride was added, and after 2 hoursand after 5 hours 0.9 mg of sodium hydride was added, while the reactionwas conducted for 7 hours under stirring.

Then, the reaction solution was poured into 100 ml of 0.1 M sodiumphosphate buffer pH 5.5 and extracted with 100 ml of ethyl acetate. Theethyl acetate extract was concentrated in vacuo, charged onto a silicagel column (Kieselgel 60, Merck, 1.5φ×30 cm) and eluted withchloroform-methanol (100:1). The fraction containing the desired productwas concentrated in vacuo to give 19 mg of a residue. Then, theresulting residue was dissolved in 1 ml of ethyl acetate and stirredtogether with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 4 hours. After the reaction,the reaction solution was filtered, and the filtrate was concentrated invacuo, subjected to chromatography using a silica gel column (Kieselgel60, Merck, 1.5φ×30 cm) and eluted with chloroform-methanol (20:1). Thefraction containing the desired product was concentrated to dryness togive 11.7 mg of compound (23).

Rf: 0.43 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 595[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.67(1H, s), 8.07 (2H, d, J=7.6 Hz),7.63(1H, t, J=7.6 Hz), 7.50(2H, t, J=7.6 Hz), 6.08(1H, brd, J=3.7 Hz),5.81(1H, brd, J=4.0 Hz), 4.94(1H, t, J=4.0 Hz), 4.89(1H, m), 4.54(1H,m), 4.04(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.82 (1H, brs),2.79(1H, t, J=3.7 Hz), 2.31(1H, m), 1.94-2.12(5H, m), 1.85(1H, m),1.76(2H, m), 1.29(1H, d, J=12.5 Hz), 1.23(1H, m), 1.02(1H, m), 1.00(3H,d, J=6.7 Hz), 0.94(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 19

Preparation of8a-[[[6,7-di(benzoyloxy)tetrahydro-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (24)

In the first silica gel chromatography in Example 18, the fractioncontaining the desired compound eluted earlier than compound (23) wasconcentrated to give a residue. The resulting residue was dissolved in 1ml of ethyl acetate and stirred together with a catalytic amount of 10%palladium-carbon under a hydrogen atmosphere at room temperature for 3hours. After the reaction, the reaction solution was filtered, and thefiltrate was concentrated in vacuo, subjected to silica gel columnchromatography (Kieselgel 60, Merck, 1.5φ×30 cm) and eluted withchloroform-methanol (20:1). The fraction containing the desired productwas concentrated to dryness to give 7.0 mg of compound (24).

Rf: 0.76 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 699[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.59(1H, s), 8.15 (4H, m), 7.63(2H, m),7.50(4H, m), 6.04(1H, brd, J=3.7 Hz), 5.96(1H, brd, J=3.0 Hz), 5.34(1H,brs), 5.02(1H, t, J=3.4 Hz), 4.96(1H, m), 4.74(1H, m), 4.08(1H, d, J=9.8Hz), 3.92(1H, d, J=9.8 Hz), 2.61(1H, t, J=3.7 Hz), 2.27(1H, m),1.70-2.00(6H, m), 1.63(1H, t, J=13.6 Hz), 1.38(1H, m), 1.16(2H, m),0.97(3H, d, J=6.7 Hz), 0.91 (1H, m), 0.90(3H, d, J=6.7 Hz), 0.69(3H, d,J=7.0 Hz).

EXAMPLE 20

Preparation of8a-[[[6-(Phenylacetyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (25)

50 mg of compound (4) was dissolved in 4 ml of dry dimethylformamide andstirred together with 5.5 mg of sodium hydride under cooling with ice.After 0.5 hour, 17.7 mg of phenylacetyl chloride was added, while thereaction was conducted for 4 hours under stirring.

Then, the reaction solution was poured into 100 ml of 0.1 M sodiumphosphate buffer pH 5.5 and extracted with 100 ml of ethyl acetate. Theethyl acetate extract was concentrated in vacuo, charged onto a silicagel column (Kieselgel 60, Merck, 1.5φ×30 cm) and eluted withn-hexane-ethyl acetate (2:1). The fraction containing the desiredproduct was concentrated in vacuo to give 18.3 mg of a residue.

Then, the resulting residue was dissolved in 1.5 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 2 hours. After the reaction,the reaction solution was filtered, and the filtrate was concentrated invacuo, subjected to silica gel column chromatography (Kieselgel 60,Merck, 1.5φ×30 cm) and eluted with chloroform-methanol (20:1). Thefraction containing the desired product was concentrated to dryness togive 10.6 mg of compound (25).

Rf: 0.23 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 609[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 7.26-7.36(5H, m), 6.10(1H,brd, J=3.7 Hz), 5.72(1H, brd, J=3.0 Hz), 4.76(1H, m), 4.70(1H, t, J=3.7Hz), 4.37(1H, m), 4.00(1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.73(2H, s), 3.59(1H, brs), 2.78(1H, t, J=3.7 Hz), 2.34(1H, m),1.74-2.10(8H, m), 1.32(1H, d, J=12.5 Hz), 1.23(1H, m), 1.05(1H, m),1.04(3H, d, J=6.7 Hz), 0.97(3H, d, J=6.7 Hz), 0.80(3H, d, J=6.7 Hz).

EXAMPLE 21

Preparation of8a-[[[6-(3-phenylpropionyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (26)

50 mg of compound (4) was dissolved in 4 ml of dry dimethylformamide andstirred together with 5.5 mg of sodium hydride under cooling with ice.After 0.5 hour, 17.4 mg of cinnamoyl chloride was added, and thereaction was conducted for 4 hours under stirring. Then, the reactionsolution was poured into 100 ml of 0.1 M sodium phosphate buffer pH 5.5and extracted with 100 mg of ethyl acetate. The ethyl acetate extractwas concentrated in vacuo, charged onto a silica gel column (Kieselgel60, Merck, 1.5φ×30 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 18 mg of a residue.

Then, the resulting residue was dissolved in 1 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 2 hours. After the reaction,the reaction solution was filtered, and the filtrate was concentrated invacuo, subjected to silica gel column chromatography (Kieselgel 60,Merck, 1.5φ×30 cm) and eluted with 200 ml of chloroform-methanol (40:1)and 200 ml of chloroform-methanol (20:1) successively. The fractioncontaining the desired product was concentrated to dryness to give 3.0mg of compound (26).

Rf: 0.45 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 623[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.65(1H, s), 7.20-7.32(5H, m), 6.10(1H,brd, J=3.7 Hz), 5.72(1H, brd, J=3.0 Hz), 4.72(2H, m), 4.34(1H, m),4.03(1H, d, J=9.8 Hz), 3.94(1H, d, J=9.8 Hz), 3.57(1H, brs), 2.97(2H, t,J=7.4 Hz), 2.70-2.80(3H, m), 2.34(1H, m), 1.70-2.10(8H, m),1.20-1.30(2H, m), 1.02(3H, d, J=6.7 Hz), 1.00(1H, m), 0.97(3H, d, J=6.7Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 22

Preparation of8a-[[[6-(ethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (29)

(Step 1) Preparation of8a-[[[6-(acetyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (5)

100 mg of compound (3) was dissolved in 0.7 ml of anhydrousdichloromethane and stirred together with 70 mg of dimethylaminopyridineand 49 μl of t-butyldimethylsilyl trifluoromethanesulfonate for 1 hourunder cooling with ice. The reaction solution was concentrated in vacuo,dissolved in methanol, charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×22 cm) and eluted with n-hexane-ethyl acetate (4:1). Thefraction containing the desired product was concentrated in vacuo togive 99 mg of compound (5) as a colorless oily substance.

Rf: 0.62 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 813[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(hydroxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (6)

90 mg of compound (5) was dissolved in 2.2 ml of methanol and stirredtogether with 121 μl of 1N NaOH for 2 hours under cooling with ice.After addition of water, the reaction solution was extracted with ethylacetate, and the ethyl acetate extract was dried over anhydrous sodiumsulfate and concentrated in vacuo to dryness to give 66 mg of compound(6) as a colorless powder.

Rf: 0.38 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 793[M+Na]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.74(1H, s), 7.25-7.43(10H, m), 6.97(1H,s), 6.09(1H, brd, J=3.7 Hz), 5.75(1H, brd, J=3.0 Hz), 4.40(1H, m),4.19(1H, d, J=9.8 Hz), 4.12(1H, m), 4.01(1H, m), 3.97(1H, d, J=9.8 Hz),3.76(1H, brs), 2.80(1H, t, J=3.7 Hz), 2.45(1H, d, J=10.4 Hz), 2.23(1H,m), 1.99(1H, m), 1.82-1.95(5H, m), 1.56(2H, m), 1.25(1H, d, J=12.5 Hz),0.98(3H, d, J=6.7 Hz), 0.92(9H, s), 0.88-0.96(2H, m), 0.73(3H, d, J=6.7Hz), 0.26(3H, d, J=6.7 Hz), 0.14(3H, s), 0.13(3H, s).

(Step 3) Preparation of8a-[[[6-(ethoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (27)

48.0 mg of compound (6) was dissolved in 0.4 ml of drydimethylformamide, and about 5 mg of sodium hydride was added undercooling with ice. After 15 minutes, 19 μl of ethyl bromide was added,and the reaction solution was stirred under cooling with ice. After 50minutes, the reaction solution was brought back to room temperature andallowed to react for another 90 minutes under stirring. Then, thereaction solution was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×25 cm) and eluted with n-hexane-ethyl acetate (4:1) to give36.8 mg of compound (27) as a colorless oily substance.

Rf: 0.57 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 799[M+H]⁺.

(Step 4) Preparation of8a-[[[6-(ethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (28)

Compound (27) was mixed with in 70 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution and stirred at room temperature. After80 minutes, the reaction solution was mixed with 15 ml of ethyl acetateand washed with 10 ml of sodium phosphate buffer (10 mM, pH 5.97) andthen with 10 ml of water twice. After the washing, the ethyl acetatelayer was dried over anhydrous sodium sulfate, filtered for removal ofsolids and concentrated in vacuo to give a crude reaction product. Thereaction product was further subjected to silica gel columnchromatography (Kieselgel 60, Merck, 1.5φ×15 cm) and eluted withn-hexane-ethyl acetate (3:1) to give 21.2 mg of compound (28) as acolorless oily substance.

Rf: 0.41 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 685[M+H]⁺.

(Step 5) Preparation of Compound (29)

21.2 mg of compound (28) was dissolved in 6 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 50 minutes. The reactionsolution was filtered, and the filtrate was concentrated in vacuo. Thereaction product was dissolved in 4 ml of methanol and washed with 4 mlof n-hexane twice. The methanol layer was concentrated in vacuo to give10.0 mg of compound (29) as a colorless oily substance.

Rf: 0.40 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 519[M+H]⁺.

¹ H-NMR(δ ppm, 300 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, d, J=3.7 Hz),5.69(1H, d, J=3.7 Hz), 4.54(1H, m), 4.32(1H, m), 4.03(1H, d, J=8.3 Hz),3.95(1H, d, J=8.3 Hz), 3.53-3.79(4H, m), 2.82(1H, t, J=3.4 Hz), 2.32(1H,m), 1.79-2.13(6H, m), 1.67-1.79(2H, m), 1.14-1.27 (2H, m), 1.26(3H, t,J=7.5 Hz), 1.02(1H, m), 1.02(3H, d, J=6.8 Hz), 0.94(3H, d, J=6.8 Hz),0.78(3H, d, J=6.8 Hz).

EXAMPLE 23

Preparation of8a-[[[6-(propoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (32)

(Step 1) Preparation of8a-[[[6-(propoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid dimethylphenylmethyl ester (30)

50 mg of compound (6) was dissolved in 1.0 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 4 mg of sodium hydrideunder cooling with ice. After 30 minutes, 56.3 μl of 1-chloropropane wasadded, and the reaction solution was stirred at room temperature for 3hours. Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate(4:1). The fraction containing the desired product was concentrated todryness to give 21.6 mg of compound (30).

Rf: 0.70 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 813[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(propoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid dimethylphenylmethyl ester (31)

21 mg of compound (30) was mixed with 53.2 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution, and the reaction was conducted atroom temperature under stirring for 2 hours. The reaction solution wascharged onto a silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) andeluted with n-hexane-ethyl acetate (2:1). The fraction containing thedesired product was concentrated to dryness to give 16 mg of compound(31).

Rf: 0.32 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 699[M+H]⁺.

(Step 3) Preparation of Compound (32)

13 mg of compound (31) was dissolved in 2 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 1 hour. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) and elutedwith chloroform-methanol (20:1). The fraction containing the desiredproduct was concentrated to dryness to give 9.0 mg of compound (32).

Rf: 0.39 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 533[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.10 (1H, brd, J=4.0 Hz),5.70(1H, brd, J=2.8 Hz), 4.55(1H, m), 4.31(1H, m), 4.03(1H, d, J=9.8Hz), 3.98(1H, d, J=9.8 Hz), 3.74(1H, d, J=0.8 Hz), 3.72(1H, t, J=3.7Hz), 3.60(1H, m), 3.49(1H, m), 2.81(1H, t, J=4.0 Hz), 2.33 (1H, m),1.90-2.10(5H, m), 1.87(1H, m), 1.76(2H, m), 1.66(2H, m), 1.29(1H, d,J=13.1 Hz), 1.22(1H, m), 1.02(3H, d, J=6.8 Hz), 1.00(1H, m), 0.95(3H, t,J=7.3 Hz), 0.95(3H, d, J=6.8 Hz), 0.79(3H, d, J=6.8 Hz).

EXAMPLE 24

Preparation of8a-[[[6-(isopropoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (35)

(Step 1) Preparation of8a-[[[6-(isopropoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (33)

50 mg of compound (6) was dissolved in 1.0 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 3 mg of sodium hydrideunder cooling with ice. After 30 minutes, 12.2 μl of 2-bromopropane wasadded, and the reaction solution was stirred at room temperature. Thereaction solution was allowed to react for 63 hours, while about 3 mg ofsodium hydride and 12.2 μl of 2-bromopropane were added after 40 hoursand further after 46 hours. Then, the reaction solution was charged ontoa silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) and eluted withn-hexane-ethyl acetate (2:1). The fraction containing the desiredproduct was concentrated to dryness to give 13.6 mg of compound (33).

Rf: 0.78 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 835[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(isopropoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (34)

13 mg of compound (33) was mixed with 200 μl of tetrahydrofuran and 14.8μl of 1M tetrabutylammonium fluoride-tetrahydrofuran solution, and thereaction was conducted at room temperature under stirring for 15 hours.The reaction solution was charged onto a silica gel column (Kieselgel60, Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated to dryness togive 9.0 mg of compound (34).

Rf: 0.30 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 699[M+H]⁺.

(Step 3) Preparation of Compound (35)

9 mg of compound (34) was dissolved in 2 ml of ethyl acetate and allowedto react in the presence of a catalytic amount of 10% palladium-carbonunder stirring under a hydrogen atmosphere at room temperature for 2hours. The reaction solution was filtered, and the filtrate wasconcentrated in vacuo. The reaction product was charged onto a silicagel column (Kieselgel 60, Merck, 1.0φ×30 cm) and eluted withchloroform-methanol (30:1). The fraction containing the desired productwas concentrated to dryness to give 6.8 mg of compound (35).

Rf: 0.39 (Kieselgel 60F254, Merck, chloroform-methanol; 50:1).

FAB-MS(m/z): 533[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.11 (1H, brd, J=3.7 Hz),5.70(1H, brd, J=4.0 Hz), 4.51(1H, m), 4.26(1H, m), 4.04(1H, d, J=9.8Hz), 3.96(1H, d, J=9.8 Hz), 3.77-3.84(2H, m), 3.75(1H, brs), 2.81(1H, t,J=3.7 Hz), 2.32(1H, m), 1.93-2.12(5H, m), 1.87(1H, m), 1.76(2H, m),1.29(1H, d, J=12.5 Hz), 1.24(3H, d, J=6.3 Hz), 1.23(3H, d, J=6.3 Hz),1.23(1H, m), 1.03(3H, d, J=7.0 Hz), 1.02(1H, m), 0.95(3H, d, J=6.7 Hz),0.79 (3H, d, J=6.7 Hz).

EXAMPLE 25

Preparation of8a-[[[6-(butoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylic acid (38)

(Step 1) Preparation of8a-[[[6-(butoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (36)

32 mg of compound (6) was dissolved in 0.4 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 5 mg of sodium hydrideunder cooling with ice. After 15 minutes, 25.6 μl of butyl bromide wasadded, and the reaction solution was stirred for 50 minutes. Thereaction solution was allowed to react at room temperature further for13 hours under stirring. Then, the reaction solution was charged onto asilica gel column (Kieselgel 60, Merck, 1.0φ×22 cm) and eluted withn-hexane-ethyl acetate (4:1) to give 26 mg of compound (36) as acolorless oily substance.

Rf: 0.72 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 849[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(butoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (37)

20 mg of compound (36) was stirred together with 242 μl of drytetrahydrofuran and 36 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 1 hour. Thereaction solution was concentrated in vacuo to give the crude reactionproduct. It was charged onto a silica gel column (Kieselgel 60, Merck,1.5φ×15 cm) and eluted with n-hexane-ethyl acetate (3:1) to give 20.5 mgof compound (37) as a colorless oily substance.

Rf: 0.38 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 713[M+H]⁺.

(Step 3) Preparation of Compound (38)

20.5 mg of compound (37) was dissolved in 1 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 1 hour. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×22 cm) and elutedwith n-hexane-ethyl acetate (1:1) to give 8.8 mg of compound (38) as acolorless oily substance.

Rf: 0.39 (Kieselgel 60F254, Merck, ethyl acetate).

FAB-MS(m/z): 569[M+Na]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.11 (1H, brd, J=3.7 Hz),5.70(1H, brd, J=2.8 Hz), 4.55(1H, m), 4.30(1H, m), 4.04(1H, d, J=9.8Hz), 3.97(1H, d, J=9.8 Hz), 3.73(1H, brs), 3.72(1H, t, J=4.0 Hz),3.64(1H, m), 3.53(1H, m), 2.81(1H, t, J=3.7 Hz), 2.33(1H, m),1.94-2.12(5H, m), 1.86(1H, m), 1.76(2H, m), 1.62(2H, m), 1.40(2H, m),1.29(1H, d, J=12.5 Hz), 1.24(1H, m), 1.03(1H, m), 1.03(3H, d, J=6.7 Hz),0.96(3H, d, J=6.7 Hz), 0.94(3H, t, J=7.3 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 26

Preparation of8a-[[[6-(pentyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (41)

(Step 1) Preparation of8a-[[[6-(pentyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (39)

50 mg of compound (6) was dissolved in 0.5 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 5 mg of sodium hydrideunder cooling with ice. After 30 minutes, 78.7 μl of 1-chloropentane wasadded, and the reaction solution was stirred at room temperature for 4hours. Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate(5:1). The fraction containing the desired product was concentrated todryness to give 22.0 mg of compound (38).

Rf: 0.79 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 841[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(pentyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (40)

22 mg of compound (39) was mixed with 500 μl of tetrahydrofuran and 52.4μl of 1M tetrabutylammonium fluoride-tetrahydrofuran solution, and thereaction was conducted at room temperature under stirring for 2 hours.

The reaction solution was charged onto a silica gel column (Kieselgel60, Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate (3:1). Thefraction containing the desired product was concentrated to dryness togive 16.6 mg of compound (40).

Rf: 0.42 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 727[M+H]⁺.

(Step 3) Preparation of Compound (41)

15 mg of compound (40) was dissolved in 2 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 1 hour. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) and elutedwith chloroform-methanol (40:1). The fraction containing the desiredproduct was concentrated to dryness to give 11.5 mg of compound (41).

Rf: 0.30 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 561[M+H]⁺.

¹ H-NMR(δ ppm, 400 MHz, CDCl₃):9.67(1H, s), 6.11(1H, dd, J=3.4, 1.0 Hz),5.70(1H, dd, J=3.4, 1.0 Hz), 4.55 (1H, m), 4.31(1H, m), 4.04(1H, d,J=9.8 Hz), 3.97(1H, d, J=9.8 Hz), 3.71-3.74(2H, m), 3.63(1H, m),3.52(1H, m), 2.81(1H, t, J=3.4 Hz), 2.33(1H, m), 1.94-2.11(5H, m),1.86(1H, m), 1.72-1.81(2H, m), 1.64(2H, m), 1.18-1.38(6H, m), 1.02(3H,d, J=6.8 Hz), 1.01(1H, m), 0.95(3H, d, J=6.3 Hz), 0.91(3H, t, J=6.8 Hz),0.79(3H, d, J=6.8 Hz).

EXAMPLE 27

Preparation of8a-[[[6-(isopentyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (44)

(Step 1) Preparation of8a-[[[6-(3-methyl-2-butenyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (42)

50 mg of compound (6) was dissolved in 0.4 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 5 mg of sodium hydrideunder cooling with ice. After 15 minutes, 37.4 ml of4-bromo-2-methyl2-butene was added, and the reaction solution wasstirred under cooling with ice for 50 minutes. The reaction solution wasallowed to react at room temperature for another 20 minutes understirring. Then, the reaction solution was charged onto a silica gelcolumn (Kieselgel 60, Merck, 1.5φ×19 cm) and eluted with n-hexane-ethylacetate (10:1) to give 34.4 mg of compound (42) as a colorless oilysubstance.

Rf: 0.72 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 861[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(3-methyl-2-butenyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (43)

31 mg of compound (42) was stirred together with 1.23 ml of drytetrahydrofuran and 55.3 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 3 hours. Thereaction solution was concentrated in vacuo to give the crude reactionproduct. It was charged onto a silica gel column (Kieselgel 60, Merck,1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (4:1) to give 31.7 mgof compound (43) as a colorless oily substance.

Rf: 0.36 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 725[M+H]⁺.

(Step 3) Preparation of Compound (44)

30 mg of compound (43) was dissolved in 4.3 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 50 minutes. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×21 cm) and elutedwith n-hexane-ethyl acetate (3:1) to give 16.5 mg of compound (44) as acolorless oily substance.

Rf: 0.12 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 561[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.11 (1H, brd, J=3.7 Hz),5.70(1H, brd, J=3.0 Hz), 4.56(1H, m), 4.30(1H, m), 4.02(1H, d, J=9.8Hz), 3.98(1H, d, J=9.8 Hz), 3.73(1H, brs), 3.72(1H, t, J=4.3 Hz),3.66(1H, m), 3.55(1H, m), 2.81(1H, t, J=3.7 Hz), 2.32(1H, m),1.93-2.10(5H, m), 1.86(1H, m), 1.75(3H, m), 1.53(2H, m), 1.29(1H, d,J=12.5 Hz), 1.23(1H, m), 1.03(3H, d, J=6.7 Hz), 1.02(1H, m), 0.95(3H, d,J=6.7 Hz), 0.92(6H, d, J=6.4 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 28

Preparation of8a-[[[6-(hexyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (47)

(Step 1) Preparation of8a-[[[6-(hexyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (45)

100.0 mg of compound (6) was dissolved in 0.8 ml of drydimethylformamide under a nitrogen atmosphere and mixed with about 10 mgof sodium hydride under cooling with ice. After 15 minutes, 44 μl ofhexyl chloride was added, and the reaction solution was stirred undercooling with ice for 2 hours. The reaction solution was allowed to reactat room temperature for another 90 minutes under stirring. Then, thereaction solution was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×21 cm) and eluted with n-hexane-ethyl acetate (4:1) to give18.7 mg of compound (45) as a colorless oily substance.

Rf: 0.77 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 877[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(hexyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (46)

18.7 mg of compound (45) was stirred together with 218 μl of drytetrahydrofuran and 32.7 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 4 hours. Thereaction solution was concentrated in vacuo to give the crude reactionproduct. It was charged onto a silica gel column (Kieselgel 60, Merck,1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (8:1) to give 16.3 mgof compound (46) as a colorless oily substance.

Rf: 0.43 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 741[M+H]⁺.

(Step 3) Preparation of Compound (47)

14.1 mg of compound (46) was dissolved in 1.9 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 3 hours. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.5φ×15 cm) and elutedwith n-hexane-ethyl acetate (3:1) to give 6.0 mg of compound (47) as acolorless oily substance.

Rf: 0.37 (Kieselgel 60F254, Merck, hexane-ethyl acetate; 1:1).

FAB-MS(m/z): 575[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.69(1H, brd, J=3.0 Hz), 4.55(1H, m), 4.30(1H, m), 4.06(1H, d, J=9.8Hz), 3.96(1H, d, J=9.8 Hz), 3.73(1H, d, J=1.5 Hz), 3.72(1H, t, J=3.7Hz), 3.63(1H, m), 3.52(1H, m), 2.79(1H, t, J=3.7 Hz), 2.33 (1H, m),1.94-2.12(5H, m), 1.85(1H, m), 1.78(2H, m), 1.63(2H, m), 1.20-1.40(8H,m), 1.04(1H, m), 1.02(3H, d, J=6.7 Hz), 0.96(3H, d, J=6.7 Hz), 0.90(3H,t, J=7.3 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 29

Preparation of8a-[[[6-(decyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (50)

(Step 1) Preparation of8a-[[[6-(decyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (48)

62.8 mg of compound (6) was dissolved in 0.4 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 7 mg of sodium hydrideunder cooling with ice. After 15 minutes, 80 μl of decyl bromide wasadded, and the reaction solution was allowed to react at roomtemperature under stirring for another 18 hours. Then, the reactionsolution was charged onto a silica gel column (Kieselgel 60, Merck,1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (4:1) to give 85.7 mgof compound (48) as a colorless oily substance.

Rf: 0.84 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 933[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(decyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (49)

85.7 mg of compound (48) was stirred together with 120 μl of 1Mtetrabutylammonium fluoride-tetrahydrofuran solution at room temperaturefor 45 minutes.

After addition of 50 ml of ethyl acetate, the reaction solution waswashed with 40 ml of sodium phosphate buffer (10 mM, pH 5.97) and then40 ml of water twice. The washed ethyl acetate layer was dried overanhydrous sodium sulfate and filtered. The resulting ethyl acetate layerwas concentrated in vacuo to give the crude reaction product.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×14 cm) and eluted with n-hexane-ethyl acetate (4:1) to give44.3 mg of compound (49) as a colorless oily substance.

Rf: 0.66 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 797[M+H]⁺.

(Step 3) Preparation of Compound (50)

44.3 mg of compound (49) was dissolved in 5 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 70 minutes. The reaction solution was filtered, and thefiltrate was concentrated in vacuo to give the crude reaction product.The crude reaction product was charged onto a silica gel column(Kieselgel 60, Merck, 1.5φ×15 cm) and eluted with 150 ml ofn-hexane-ethyl acetate (2:1), 200 ml of n-hexane-ethyl acetate (1:3) andfinally chloroform-methanol (10:1) to give 19.8 mg of compound (50) as acolorless oily substance.

Rf: 0.15 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 631[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.65(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.70(1H, brd, J=4.0 Hz), 4.55(1H, m), 4.31(1H, m), 4.04(1H, d, J=9.8Hz), 4.97(1H, d, J=9.8 Hz), 3.74(1H, brs), 3.72(1H, t, J=3.7 Hz),3.63(1H, m), 3.52(1H, m), 2.79(1H, brt, J=4.0 Hz), 2.34(1H, m),1.84-2.12(6H, m), 1.75(2H, m), 1.63(2H, m), 1.18-1.38(16H, m), 1.02(3H,d, J=6.7 Hz), 1.01(1H, m), 0.96 (3H, d, J=6.7 Hz), 0.88(3H, t, J=7.3Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 30

Preparation of8a-[[[6-(cetyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (53)

(Step 1) Preparation of8a-[[[6-(cetyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (51)

115 mg of compound (6) was dissolved in 1.5 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with 13.8 mg of sodium hydrideunder cooling with ice. After 30 minutes, 225 μl of cetyl chloride wasadded, and the reaction solution was stirred under cooling with ice.After 30 minutes, the reaction solution was brought back to roomtemperature and allowed to react for another 90 minutes under stirring.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.5φ×22 cm) and eluted with n-hexane-ethyl acetate(8:1) to give 43.3 mg of compound (51) as a colorless oily substance.

Rf: 0.80 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 1017[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(cetyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (52)

43.3 mg of compound (51) was stirred together with 435 μl of drytetrahydrofuran and 65.3 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 1 hour. Thereaction solution was concentrated in vacuo to give the crude product.The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (8:1) to give22.5 mg of compound (52) as a colorless oily substance.

Rf: 0.53 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 903[M+Na]⁺.

(Step 3) Preparation of Compound (53)

25.0 mg of compound (52) was dissolved in 2.89 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 1 hour. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×21 cm) and elutedwith n-hexane-chloroform (2:1) to give 12.3 mg of compound (53) as acolorless oily substance.

Rf: 0.15 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 737[M+Na]⁺.

¹ H-NMR(δ ppm, 400 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, dd, J=3.4, 1.0Hz), 5.70(1H, dd, J=3.4, 1.5 Hz), 4.55(1H, m), 4.30(1H, m), 4.03(1H, d,J=9.8 Hz), 3.96 (1H, d, J=9.8 Hz), 3.70-3.74(2H, m), 3.63(1H, m),3.52(1H, m), 2.81(1H, t, J=3.4 Hz), 2.32(1H, m), 1.92-2.12(5H, m),1.87(1H, m), 1.72-1.80(2H, m), 1.62 (2H, m), 1.17-1.39(28H, m), 1.02(3H,d, J=6.8 Hz), 1.00(1H, m), 0.95(3H, d, J=6.8 Hz), 0.88(3H, t, J=6.8 Hz),0.78(3H, d, J=6.8 Hz).

EXAMPLE 31

Preparation of8a-[[[6-(2-(N-piperidino)ethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (56)

(Step 1) Preparation of8a-[[[6-(2-(N-piperidino)ethoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (54)

67.6 mg of compound (6) was dissolved in 0.2 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 20 mg of sodium hydrideunder cooling with ice. After 15 minutes, 57 mg of1-(2-chloroethyl)piperidine hydrochloride suspended in 1 ml ofdimethylformamide was added, and the reaction solution was allowed toreact at room temperature under stirring for 66 hours.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×30 cm) and eluted with 225 ml ofn-hexane-ethyl acetate (2:1) and chloroform-methanol (10:1). Thefraction containing the desired product was concentrated in vacuo. Theconcentrate was dissolved in 100 ml of ethyl acetate and washed with 100ml of water four times. The ethyl acetate layer was concentrated to give33.0 mg of compound (54) as a colorless oily substance.

Rf: 0.23 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 882[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(2-(N-piperidino)ethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (55)

33.0 mg of compound (54) was stirred together with 60 μl of 1Mtetrabutylammonium fluoride-tetrahydrofuran solution at room temperaturefor 45 minutes.

The reaction solution was charged onto a silica gel column (Kieselgel60, Merck, 1.5φ×15 cm) and eluted with 25 ml of chloroform and thenchloroform-methanol (5:1) to give 26.7 mg of compound (55) as acolorless oily substance.

Rf: 0.28 (Kieselgel 60F254, Merck, chloroform-methanol; 5:1).

FAB-MS(m/z): 768[M+H]⁺.

(Step 3) Preparation of Compound (56)

26.7 mg of compound (55) was dissolved in 5 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature. After 5 hours, 3 ml of aceticacid was added. Then, 4 hours later, 3 ml of methanol was further added,and the reaction was conducted for 1 hour. The reaction solution wasfiltered, mixed with 5 ml of sodium phosphate buffer (10 mM, pH 5.97)and washed with 10 ml of ethyl acetate. The ethyl acetate layer waswashed with 5 ml of water twice. All the lower layers were combined andconcentrated to 10 ml in vacuo for removal of ethyl acetate byevaporation. The concentrate was charged onto Diaion HP-20 (MitsubishiChemical, volume 3 ml), washed with 20 ml of water and eluted with 15 mlof methanol. The methanolic eluate was concentrated in vacuo to give 7.5mg of compound (56) as a colorless solid.

Rf: 0.13 (Kieselgel 60F254, Merck, chloroform-methanol; 5:1).

FAB-MS(m/z): 602[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.81 (1H, s), 5.94 (1H, brd, J=3.7 Hz),5.64(1H, brd, J=3.0 Hz), 4.88(1H, m), 4.21(1H, m), 4.16(1H, m), 4.00(1H,d, J=9.8 Hz), 3.92(1H, d, J=9.8 Hz), 3.78(2H, m), 3.63(1H, brs),3.24(1H, m), 2.76(1H, m), 2.71(1H, t, J=3.7 Hz), 2.40 (1H, m), 2.23(1H,m), 1.50-2.10(13H, m), 1.20(2H, m), 1.00(1H, m), 0.98(3H, d, J=6.7 Hz),0.95(3H, d, J=6.7 Hz), 0.76(3H, d, J=6.7 Hz).

EXAMPLE 32

Preparation of8a-[[[6-(3-phenoxypropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (59)

(Step 1) Preparation of8a-[[[6-(3-phenoxypropyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (57)

49.5 mg of compound (6) was dissolved in 0.4 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 7 mg of sodium hydrideunder cooling with ice. After 15 minutes, 40 μl of 3-phenoxypropylbromide was added, and the reaction was conducted for 66 hours understirring.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.5φ×25 cm) and eluted with n-hexane-ethyl acetate(4:1). The fraction containing the desired product was concentrated invacuo to give 34.5 mg of compound (57) as a colorless oily substance.

Rf: 0.51 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 905[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(3-phenoxypropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (58)

34.5 mg of compound (57) was dissolved in 0.1 ml of dry tetrahydrofuranand stirred together with 60 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 115 minutes.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×13 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 13.0 mg of compound (58) as a colorless oily substance.

Rf: 0.26 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 791[M+H]⁺.

(Step 3) Preparation of Compound (59)

13.0 mg of compound (58) was dissolved in 2 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature. After 30 minutes, the reactionsolution was filtered and concentrated in vacuo. The reaction productwas dissolved in 2 ml of methanol and washed with 2 ml of n-hexanetwice, and the lower layer was concentrated in vacuo to give 6.6 mg ofcompound (59) as a colorless solid.

Rf: 0.49 (Kieselgel 60F254, Merck, chloroform-methanol; 10:1).

FAB-MS(m/z): 625[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 7.28 (2H, t, J=7.6 Hz),6.95(1H, t, J=7.6 Hz), 6.90(2H, d, J=7.6 Hz), 6.06(1H, brd, 3.7 Hz),5.69(1H, brd, J=3.0 Hz), 4.55(1H, m), 4.31(1H, m), 4.09(2H, t, J=6.1Hz), 4.03 (1H, d, J=9.8 Hz), 3.96(1H, d, J=9.8 Hz), 3.82(1H, m),3.75(2H, m), 3.72(1H, brs), 2.75(1H, t, J=3.7 Hz), 2.33 (1H, m),2.11(2H, m), 1.83-2.10(6H, m), 1.74(2H, m), 1.26(1H, d, J=12.5 Hz),1.22(1H, m), 1.01(3H, d, J=6.7 Hz), 1.01(1H, m), 0.94(3H, d, J=6.7 Hz),0.78(3H, d, J=6.7 Hz).

EXAMPLE 33

Preparation of8a-[[[6-(3-(N-1H-pyrrolo)propyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (62)

(Step 1) Preparation of8a-[[[6-(3-(N-1H-pyrrolo)propyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (60)

50.0 mg of compound (6) was dissolved in 0.4 ml of dry dimethylformamideunder a nitrogen atmosphere and mixed with about 8 mg of sodium hydrideunder cooling with ice. After 20 minutes, 40 μl of1-(3-bromopropyl)pyrrole was added, and the reaction was conducted atroom temperature under stirring for 110 minutes.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.5φ×15 cm) and eluted with n-hexane-ethyl acetate(4:1). The fraction containing the desired product was concentrated invacuo to give 47.2 mg of compound (60) as a colorless oily substance.

Rf: 0.40 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 878[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(3-(N-1H-pyrrolo)propyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (61)

42.7 mg of compound (60) was dissolved in 0.2 ml of dry tetrahydrofuranand stirred together with 80 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 70 minutes.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×15 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 36.0 mg of compound (61) as a colorless oily substance.

Rf: 0.29 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 764[M+H]⁺.

(Step 3) Preparation of Compound (62)

36.0 mg of compound (61) was dissolved in a liquid mixture of 3 ml ofethyl acetate and 3 ml of methanol and stirred together with a catalyticamount of 10% palladium-carbon under a hydrogen atmosphere at roomtemperature. After 40 minutes, the reaction solution was filtered andconcentrated in vacuo. The reaction product was dissolved in 5 ml ofmethanol and washed with 5 ml of n-hexane twice, and the lower layer wasconcentrated in vacuo to give 24.2 mg of compound (62) as a colorlesssolid.

Rf: 0.22 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 1:1).

FAB-MS(m/z): 598[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.68(1H, s), 6.67 (2H, t, J=2.1 Hz),6.14(2H, t, J=2.1 Hz), 6.11(1H, brd, J=3.7 Hz), 5.71(1H, brd, J=3.0 Hz),4.54(1H, m), 4.28 (1H, m), 4.00-4.06(4H, m), 3.74(1H, brs), 3.69(1H, t,J=4.0 Hz), 3.55(1H, m), 3.44(1H, m), 2.83(1H, t, J=3.7 Hz), 2.34(1H, m),1.90-2.10(7H, m), 1.87(1H, m), 1.76(2H, m), 1.20-1.30(2H, m), 1.03(3H,d, J=6.7 Hz), 1.02(1H, m), 0.97(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 34

Preparation of8a-[[[6-(cyanopropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (65)

(Step 1) Preparation of8a-[[[6-(cyanopropyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (63)

150.0 mg of compound (6) was dissolved in 0.4 ml of drydimethylformamide under a nitrogen atmosphere and mixed with 14 mg ofsodium hydride under cooling with ice. After 30 minutes, 100 μl of4-bromobutyronitrile was added, and the reaction was conducted undercooling with ice under stirring for 2.5 hours.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×28 cm) and eluted with n-hexane-ethyl acetate(4:1). The fraction containing the desired product was concentrated invacuo to give 27.3 mg of compound (63) as a colorless oily substance.

Rf: 0.71 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 1:1).

FAB-MS(m/z): 860[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(cyanopropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (64)

24 mg of compound (63) was dissolved in 1.0 ml of dry tetrahydrofuranand stirred together with 44 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 1.5 hours.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×21 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 22.4 mg of compound (64) as a colorless oily substance.

Rf: 0.36 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 724[M+H]⁺.

(Step 3) Preparation of Compound (65)

20 mg of compound (64) was dissolved in 3 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 1.5 hours. The reactionsolution was filtered and concentrated in vacuo. The reaction productwas dissolved in 10 ml of methanol and washed with 10 ml of n-hexanefour times, and the lower layer was concentrated in vacuo to give 4.2 mgof compound (65) as a colorless solid.

Rf: 0.15 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 558[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.64(1H, s), 6.09 (1H, brd, J=3.7 Hz),5.72(1H, brd, J=3.0 Hz), 4.58(1H, m), 4.32(1H, m), 4.05(1H, d, J=9.8Hz), 3.95(1H, d, J=9.8 Hz), 3.77(1H, t, J=4.0 Hz), 3.72(1H, m), 3.71(1H,brs), 3.68(1H, m), 2.74(1H, brs), 2.52(2H, t, J=7.3 Hz), 2.36(1H, m),1.70-2.10(10H, m), 1.18-1.30(2H, m), 1.02(3H, d, J=6.7 Hz), 1.01(1H, m),0.97(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 35

Preparation of8a-[[[6-(ethoxycarbonylmethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (68)

(Step 1) Preparation of8a-[[[6-(ethoxycarbonylmethoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (66)

50.0 mg of compound (6) was dissolved in 0.65 ml of drydimethylformamide under a nitrogen atmosphere and mixed with 4.6 mg ofsodium hydride under cooling with ice. After 30 minutes, 36 μl of ethylbromoacetate was added, and the reaction was conducted under coolingwith ice under stirring for 3.0 hours.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×28 cm) and eluted with n-hexane-ethyl acetate(8:1). The fraction containing the desired product was concentrated invacuo to give 27.3 mg of compound (66) as a colorless oily substance.

Rf: 0.50 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 879[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(ethoxycarbonylmethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (67)

21 mg of compound (66) was dissolved in 0.8 ml of dry tetrahydrofuranand stirred together with 36 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 1.5 hours.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 20.1 mg of compound (67) as a colorless oily substance.

Rf: 0.22 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 743[M+H]⁺.

(Step 3) Preparation of Compound (68)

22.4 mg of compound (67) was dissolved in 3 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 1.5 hours. The reactionsolution was filtered and concentrated in vacuo. The reaction productwas dissolved in 5 ml of methanol and washed with 5 ml of n-hexanetwice, and the lower layer was concentrated in vacuo to give 6.8 mg ofcompound (68) as a colorless solid.

Rf: 0.15 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 577[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.67(1H, s), 6.10 (1H, brd, J=3.7 Hz),5.70(1H, brd, J=3.0 Hz), 4.62(1H, m), 4.37(1H, m), 4.30(1H, d, J=16.8Hz), 4.23(2H, t, J=7.0 Hz), 4.19(1H, d, J=16.8 Hz), 4.03(1H, d, J=9.8Hz), 3.97(1H, d, J=9.8 Hz), 3.86(2H, m), 2.82(1H, t, J=3.7 Hz), 2.32(1H,m), 1.84-2.10(6H, m), 1.75(2H, m), 1.30(3H, t, J=7.0 Hz), 1.18-1.30(2H,m), 1.02(3H, d, J=6.7 Hz), 1.02(1H, m), 0.94(3H, d, J=6.7 Hz), 0.78(3H,d, J=6.7 Hz).

EXAMPLE 36

Preparation of8a-[[[6-(aminocarbonylmethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (71)

(Step 1) Preparation of8a-[[[6-(aminocarbonylmethoxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (69)

50.0 mg of compound (6) was dissolved in 0.65 ml of drydimethylformamide under a nitrogen atmosphere and mixed with 4.6 mg ofsodium hydride under cooling with ice. After 30 minutes, 43 μl ofbromoacetamide was added, and the reaction was conducted under coolingwith ice under stirring for 3 hours.

Then, the reaction solution was charged onto a silica 15 gel column(Kieselgel 60, Merck, 1.0φ×28 cm) and eluted with n-hexane-ethyl acetate(1:1) and then ethyl acetate. The fraction containing the desiredproduct was concentrated in vacuo to give 32.5 mg of compound (69) as acolorless oily substance.

Rf: 0.13 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 828[M+H]⁺.

(Step 2) Preparation of8a-[[[6-(aminocarbonylmethoxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (70)

21 mg of compound (69) was dissolved in 0.8 ml of dry tetrahydrofuranand stirred together with 60 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 3 hours.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×20 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated in vacuo togive 17 mg of compound (70) as a colorless oily substance.

Rf: 0.30 (Kieselgel 60F254, Merck, ethyl acetate).

FAB-MS(m/z): 714[M+H]⁺.

(Step 3) Preparation of Compound (71)

14 mg of compound (70) was dissolved in 3 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 1.5 hours. The reactionsolution was filtered and concentrated in vacuo. The reaction productwas dissolved in 10 ml of methanol and washed with 10 ml of n-hexanefour times, and the lower layer was concentrated in vacuo to give 8.6 mgof compound (71) as a colorless solid.

Rf: 0.15 (Kieselgel 60F254, Merck, ethyl acetate).

FAB-MS(m/z): 548[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 6.81 (1H, brs), 6.17(1H,brs), 6.10(1H, brd, J=3.7 Hz), 5.74 (1H, brd, J=3.0 Hz), 4.60(1H, m),4.34(1H, m), 4.18(1H, d, J=15.6 Hz), 4.11(1H, d, J=15.6 Hz), 4.02(1H, d,J=9.8 Hz), 3.98(1H, d, J=9.8 Hz), 3.85(1H, t, J=4.0 Hz), 3.74(1H, brs),2.81(1H, t, J=3.7 Hz), 2.34(1H, m), 1.93-2.12(5H, m), 1.86(1H, m),1.75(2H, m), 1.31(1H, d, J=12.5 Hz), 1.23(1H, m), 1.03(3H, d, J=6.7 Hz),1.03 (1H, m), 0.96(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 37

Preparation of8a-[[[6-(hydroxypropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (74)

(Step 1) Preparation of8a-[[[6-(benzyloxypropyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (72)

50.0 mg of compound (6) was dissolved in 0.65 ml of drydimethylformamide under a nitrogen atmosphere and mixed with 4.6 mg ofsodium hydride under cooling with ice. After 30 minutes, 57 μl of benzyl3-bromopropyl ether was added, and the reaction was conducted undercooling with ice under stirring for 30 minutes.

Then, the reaction solution was charged onto a silica gel column(Kieselgel 60, Merck, 1.0φ×20 cm) and eluted with n-hexane-ethyl acetate(10:1) and then n-hexane-ethyl acetate (3:1). The fraction containingthe desired product was concentrated in vacuo to give 50.7 mg ofcompound (72) as a colorless oily substance.

Rf: 0.62 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 941[M+Na]⁺.

(Step 2) Preparation of8a-[[[6-(benzyloxypropyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (73)

50 mg of compound (72) was dissolved in 1.8 ml of dry tetrahydrofuranand stirred together with 81 μl of 1M tetrabutylammoniumfluoride-tetrahydrofuran solution at room temperature for 2.5 hours.

The reaction product was charged onto a silica gel column (Kieselgel 60,Merck, 1.5φ×20 cm) and eluted with n-hexane-ethyl acetate (5:1). Thefraction containing the desired product was concentrated in vacuo togive 42.3 mg of compound (73) as a colorless oily substance.

Rf: 0.26 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 805[M+H]⁺.

(Step 3) Preparation of Compound (74)

38.3 mg of compound (73) was dissolved in 0.9 ml of ethyl acetate andstirred together with a catalytic amount of 10% palladium-carbon under ahydrogen atmosphere at room temperature for 2.5 hours. The reactionsolution was filtered and concentrated in vacuo. The reaction productwas dissolved in 10 ml of methanol and washed with 15 ml of n-hexanefour times, and the lower layer was concentrated in vacuo to give 23.5mg of compound (74) as a colorless solid.

Rf: 0.36 (Kieselgel 60F254, Merck, ethyl acetate).

FAB-MS(m/z): 549[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.66(1H, s), 6.15 (1H, brd, J=3.7 Hz),5.72(1H, brd, J=3.0 Hz), 4.60(1H, m), 4.32(1H, m), 4.00(2H, s),3.70-3.83(6H, m), 2.81 (1H, t, J=3.7 Hz), 2.33(1H, m), 1.93-2.12(5H, m),1.74-1.90(5H, m), 1.30(1H, d, J=12.5 Hz), 1.23(1H, m), 1.05(1H, m),1.03(3H, d, J=6.7 Hz), 0.97(3H, d, J=6.7 Hz), 0.79(3H, d, J=6.7 Hz).

EXAMPLE 38

Preparation of8a-[[[6-(benzyloxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (76)

(Step 1) Preparation of8a-[[[6-(benzyloxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (75)

1.12 g of compound (3) was dissolved in 10 ml of dichloromethane andstirred together with 42.3 mg of sodium hydride at room temperature.274.4 mg of benzyl chloride and 21.1 mg of sodium hydride were addedafter 0.5 hour and after 2 hours, respectively, and the reactionsolution was stirred for another 2 hours.

Then, the reaction solution was mixed with 5 ml of methanol and 1.5 mlof 1N sodium hydroxide, then stirred for 3 hours and poured into 500 mlof 0.1M sodium phosphate buffer pH 5.5 and extracted with 500 ml ofethyl acetate. The ethyl acetate extract was concentrated in vacuo,charged onto a silica gel column (Kieselgel 60, Merck, 3.0φ×40 cm) andeluted with chloroform-methanol (200:1). The fraction containing thedesired product was concentrated in vacuo to give 320 mg of a residue.

Then, the residue was charged onto a silica gel column (Kieselgel 60,Merck, 2.5φ×40 cm) and eluted with chloroform-n-hexane-ethyl acetate(1:2:1). The fraction containing the desired product was concentrated todryness to give 91 mg of compound (75).

Rf: 0.41 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 747[M+H]⁺.

(Step 2) Preparation of Compound (76)

15 mg of compound (75) was dissolved in 1 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 1 hour. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) and elutedwith chloroform-methanol (20:1). The fraction containing the desiredproduct was concentrated to dryness to give 10.0 mg of compound (76).

Rf: 0.26 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 581[M+H]⁺.

¹ H-NMR(δ ppm, 400 MHz, CDCl₃):9.68(1H, s), 7.31-7.38(5H, m), 6.12(1H,dd, J=3.4, 1.5 Hz), 5.66(1H, dd, J=3.4, 1.5 Hz), 4.74(1H, d, J=11.7 Hz),4.62(1H, d, J=11.7 Hz), 4.53(1H, m), 4.17(1H, m), 4.04(1H, d, J=9.8 Hz),3.98(1H, d, J=9.8 Hz), 3.76-3.78(2H, m), 2.83(1H, t, J=3.4 Hz), 2.33(1H,m), 1.94-2.10(5H, m), 1.87(1H, m), 1.72-1.80(2H, m), 1.18-1.32(2H, m),1.03(3H, d, J=6.8 Hz), 1.01(1H, m), 0.96(3H, d, J=6.8 Hz), 0.78(3H, d,J=6.8 Hz).

EXAMPLE 39

Preparation of8a-[[[6-(benzylaminocarboxy)tetrahydro-7-hydroxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid (79)

(Step 1) Preparation of8a-[[[6-(benzylaminocarboxy)tetrahydro-7-t-butyldimethylsilyloxy-2,5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (77)

50 mg of compound (6) was dissolved in dry toluene under a nitrogenatmosphere and stirred together with 9.6 μl of benzyl isocyanate and acatalytic amount of dibutyltin diacetate at room temperature for 20hours. After the reaction, the insolubles were filtered off, and thefiltrate was concentrated in vacuo to give a residue.

Then, the residue was charged onto a silica gel column (Kieselgel 60,Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate (4:1). Thefraction containing the desired product was concentrated to dryness togive 40 mg of compound (77).

Rf: 0.53 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 2:1).

FAB-MS(m/z): 904[M+H]⁺.

(Step 2) Preparation of 8a-[[[6-(benzylaminocarboxy)tetrahydro-7-hydroxy-2, 5-methanofuro[2,3-d]-1,3-dioxol-2-yl]oxy]methyl]-4-formyl-4,4a,5,6,7,7a,8,8a-octahydro-7-methyl-3-(1-methylethyl)-1,4-methano-s-indacene-3a(1H)-carboxylicacid diphenylmethyl ester (78)

40 mg of compound (77) was mixed with 1.0 ml of tetrahydrofuran and 66.4μl of 1M tetrabutylammonium fluoride-tetrahydrofuran solution, and thereaction was conducted at room temperature under stirring for 1 hour.

The reaction solution was charged onto a silica gel column (Kieselgel60, Merck, 1.0φ×30 cm) and eluted with n-hexane-ethyl acetate (2:1). Thefraction containing the desired product was concentrated to dryness togive 32 mg of compound (78).

Rf: 0.55 (Kieselgel 60F254, Merck, n-hexane-ethyl acetate; 1:1).

FAB-MS(m/z): 790[M+H]⁺.

(Step 3) Preparation of Compound (79)

27 mg of compound (78) was dissolved in 2 ml of ethyl acetate andallowed to react in the presence of a catalytic amount of 10%palladium-carbon under stirring under a hydrogen atmosphere at roomtemperature for 3 hours. The reaction solution was filtered, and thefiltrate was concentrated in vacuo. The reaction product was chargedonto a silica gel column (Kieselgel 60, Merck, 1.0φ×30 cm) and elutedwith chloroform-methanol (20:1). The fraction containing the desiredproduct was concentrated to dryness to give 18.4 mg of compound (79).

Rf: 0.36 (Kieselgel 60F254, Merck, chloroform-methanol; 20:1).

FAB-MS(m/z): 624[M+H]⁺.

¹ H-NMR(δ ppm, 500 MHz, CDCl₃):9.69(1H, s), 7.26-7.36(5H, m), 6.09(1H,brd, J=3.7 Hz), 5.74(1H, brd, J=2.2 Hz), 5.24(1H, brt, J=5.8 Hz),4.77(1H, t, J=3.7 Hz), 4.72(1H, m), 4.35-4.43(3H, m), 4.02(1H, d, J=9.8Hz), 3.95(1H, d, J=9.8 Hz), 3.66(1H, brs), 2.80(1H, t, J=3.7 Hz),2.32(1H, m), 1.90-2.12(5H, m), 1.87(1H, m), 1.74(2H, m), 1.29(1H, d,J=12.8 Hz), 1.23(1H, m), 1.02(3H, d, J=6.7 Hz), 1.02(1H, m), 0.95(3H, d,J=6.7 Hz), 0.78(3H, d, J=6.7 Hz).

FORMULATION EXAMPLE 1

10 parts of compound 14, 15 parts of heavy magnesium oxide and 75 partsof lactose were uniformly mixed to obtain a powdery or fine granularpowder having a particle size of at most 350 μm. This powder was putinto capsule containers to obtain a capsule drug.

FORMULATION EXAMPLE 2

45 parts of compound 14, 15 parts of starch, 16 parts of lactose, 21parts of crystalline cellulose, 3 parts of polyvinyl alcohol and 30parts of distilled water were uniformly mixed, then pulverized,granulated and dried and then sieved to obtain granules having a sizewith a diameter of from 1410 to 177 μm.

FORMULATION EXAMPLE 3

Granules were prepared in the same manner as in Formulation Example 2,and then 96 parts of the granules were mixed with 3 parts of calciumstearate and press-molded into tablets having a diameter of 10 mm.

FORMULATION EXAMPLE 4

90 parts of granules obtained by the method of Formulation Example 2were mixed with 10 parts of crystalline cellulose and 3 parts of calciumstearate, press-molded into tablets having a diameter of 8 mm. Then, asuspension mixture of syrup gelatin and precipitated calcium carbonatewas added thereto to give sugar-coated tablets.

FORMULATION EXAMPLE 5

1 part of compound 14, 49.5 parts of Macrogol 4,000 and 49.5 parts ofMacrogol 400 were mixed and well kneaded to homogeneity to give anointment.

FORMULATION EXAMPLE 6

5 parts of compound 14, 5 parts of miconazole, 15 parts of heavymagnesium oxide and 75 parts of lactose were uniformly mixed to obtain apowdery or fine granular powder having a particle size of at most 350μm. This is powder was put into capsule containers to obtain a capsuledrug.

FORMULATION EXAMPLE 7

35 parts of compound 14, 10 parts of miconazole, 15 parts of starch, 16parts of lactose, 21 parts of crystalline cellulose, 30 parts of polydistilled water were uniformly mixed, then pulverized, granulated anddried and then sieved to obtain granules having a size with a diameterof from 1410 to 177 μm.

FORMULATION EXAMPLE 8

Granules were prepared in the same manner as in Formulation Example 7,and then 96 parts of the granules were mixed with 3 parts of calciumstearate and press-molded into tablets having a diameter of 10 mm.

FORMULATION EXAMPLE 9

90 parts of granules obtained by the method of Formulation Example 7were mixed with 10 parts of crystalline cellulose and 3 parts of calciumstearate, press-molded into tablets having a diameter of 8 mm. Then, asuspension mixture of syrup gelatin and precipitated calcium carbonatewas added thereto to give sugar-coated tablets.

FORMULATION EXAMPLE 10

0.3 part of compound 14, 0.3 part of miconazole, 2.4 parts of a nonionicsurfactant and 97 parts of physiological saline were mixed underheating, put into ampoules and sterilized to give an injection.

FORMULATION EXAMPLE 11

0.5 part of compound 14, 0.5 part of miconazole, 49.5 parts of Macrogol4,000 and 49.5 parts of Macrogol 400 were mixed and well kneaded tohomogeneity to give an ointment.

REFERENCE EXAMPLE 1

Production of BE-31405

Fungus BE-31405 strain incubated on a slant agar plate was inoculatedinto four 500 ml Erlenmeyer flasks containing 110 ml of a culture medium(pH 6.0 before sterilization) comprising 0.3% polypeptone, 0.3% glucose,1.0% wheat embryo, 0.5% gluten meal, 0.3% malt extract, 3.0% maltose,0.2% sodium chloride, 0.1% sodium nitrate, 0.1% potassiumdihydrogenphosphate, 0.05% magnesium sulfate, 0.0002% ferrous sulfate,0.00004% cupric chloride, 0.00004% manganese chloride, 0.00004% cobaltchloride, 0.00008% zinc sulfate, 0.00008% sodium borate and 0.00024%ammonium molybdate and incubated on a rotary shaker (180 rpm) at 28° C.for 72 hours. 2 ml of these cultures were inoculated into each of fifty500 ml Erlenmeyer flasks containing 110 ml of the above culture mediumand incubated on a rotary shaker (180 rpm) at 28° C. for 72 hours.

The culture solution (about 5L) thus obtained was thermally sterilizedat 90° C. for 10 minutes and filtered, and the filtrate was applied to a1.2L Diaion HP-20 adsorption column. The column was washed with 30%methanol (4L), and 3L of methanol was applied to elute the activecomponent. The methanolic eluate was concentrated in vacuo, and waterwas added to a total volume of 500 ml. It was extracted with 500 ml ofethyl acetate twice, and the resulting ethyl acetate extract wasconcentrated in vacuo to dryness. The resulting crude product wassubjected to silica gel column chromatography (inner diameter 2 cm,length 30 cm, Kieselgel 60, Merck) and eluted with 400 ml (100:1) and800 ml (50:1) of chloroform/methanol solvent mixtures successively. Thefraction containing BE-31405 was concentrated in vacuo to dryness togive 320 mg of a crude product. Then, the crude product was subjected toreversed phase HPLC (Chromatolex-OSD (100 Å-5 μm), 20Φ×250 mm, FujiDevison Chemical) using 70% aqueous methanol as the mobile phase, andthe fraction corresponding to a peak identified at about 22 minutes byUV detection at 220 nm when the flow rate was 9 ml/min was collected andconcentrated in vacuo to dryness. The resulting crude BE-31405 wassubjected to Sephadex LH20 column chromatography (inner diameter 1.5 cm,length 90 cm) using methanol as the eluent, and the fraction containingpure BE-31405 was concentrated in vacuo to dryness to give BE-31405 as awhite solid.

Production of BE-31405 using Penicillium sp. F31405-17M is shown in thefollowing Reference Examples.

In the following Reference Examples, the separation and quantificationof BE-31405 by high performance liquid chromatography (HPLC) wereconducted under the following conditions.

HPLC conditions

Column: YMC Pack ODS-A 250×4.6 mm I.D.

Column temp.: 40° C.

Mobile phase: 10% acetonitrile (containing 0.0375% trifluoroaceticacid)/80% acetonitrile (containing 0.025% trifluoroacetic acid)=55/45

Retention time: 17 minutes

Flow rate: 1.2 ml/min

Detection: 220 nm

REFERENCE EXAMPLE 2

Spores from Penicillium sp. F-31405 strain incubated on a slant agarplate containing 0.2% potato exudate powder, 1% glucose and 1.5% agar(pH 5.6 before sterilization) were suspended in 10 ml of sterilizedwater and diluted with sterilized water by factors of 10, 100, 1000,10000 and 100000. 0.2 ml of each dilute solution was spread on an agarplate and incubated at 25° C. for 4 days, and the viable colonies weretransplanted onto slant agar plates and incubated at 25° C. for 14 days.Thus, a monosporous isolate from Penicillium sp. F-31405, the F31405-17Mstrain, was obtained. The monosporous isolate F31405-17M strainincubated on slant agar plates was inoculated into a 500 ml Erlenmeyerflasks containing 110 ml of a Czapek-Dox medium (pH 6.0 beforesterilization) containing 3.6% glucose, 0.2% sodium nitrate, 0.1%dipotassium phosphate, 0.05% magnesium sulfate, 0.05% potassium chlorideand 0.001% ferrous sulfate and incubated on a rotary shaker (180 rpm) at28° C. for 72 hours. 2 ml of this culture was inoculated into a 500 mlErlenmeyer flasks containing 110 ml of a modified medium A (pH 6.0before sterilization) comprising 7.2% glucose, 0.2% sodium nitrate, 0.1%dipotassium phosphate, 0.1% magnesium sulfate, 0.05% potassium chloride,2% yeast extract and 0.5% nicotinic acid and incubated on a rotaryshaker (180 rpm) at 28° C. for 168 hours. 15 ml of the culture wasextracted with 15 ml of ethyl acetate over 30 minutes under stirring. 10ml of the ethyl acetate layer was evaporated in vacuo for removal ofethyl acetate, and the residue was dissolved in 1 ml of methanol. TheBE-31405 concentration of the methanol solution was determined by highperformance liquid chromatography.

REFERENCE EXAMPLE 3

The monosporous isolate from Penicillium sp. F-31405, the F31405-17Mstrain, incubated on a slant agar plate was inoculated into two 500 mlErlenmeyer flasks containing 110 ml of a modified medium B (pH 6.0before sterilization) comprising 7.2% glucose, 0.2% sodium nitrate, 0.1%dipotassium phosphate, 0.1% magnesium sulfate and 0.05% potassiumchloride and incubated on a rotary shaker (180 rpm) at 28° C. for 72hours. 200 ml of this culture was inoculated into a 20L fermentercontaining 10L of a modified medium C (pH 6.0 before sterilization)comprising 7.2% glucose, 0.1% magnesium sulfate, 2% yeast extract and0.2% nicotinic acid and incubated at 28° C. for 9 days with aeration (20L/min) and stirring (300 rpm). 15 ml of this culture was extracted with15 ml of ethyl acetate over 30 minutes under stirring. 10 ml of theethyl acetate layer was evaporated for removal of ethyl acetate invacuo, and the residue was dissolved in 1 ml of methanol. The BE-31405concentration of the methanol solution was determined by highperformance liquid chromatography.

INDUSTRIAL APPLICABILITY

The compounds or antifungal composition of the present invention haveexcellent antifungal activities and thus are useful as antifungalagents.

What is claimed is:
 1. A compound represented by general formula (I) ora pharmaceutically acceptable salt or ester thereof: ##STR8## whereineach of R¹ and R² is independently a hydrogen atom, a C₁ -C₁₆ alkylgroup, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ arylgroup, a C₇ -C₁₅ aralkyl group or a heterocyclic group which is notsubstituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which has one to five substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, C₁-C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxy groups, amino groups,mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆ alkylamino groups, carboxylgroups, C₁ -C₁₆ alkyloxycarbonyl groups, aminocarbonyl groups, sulfogroups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxy groups, C₇ -C₁₅ aralkyloxygroups and heterocyclic groups, or a group represented by --Y--R³ ; Y isa carbonyl group, a thiocarbonyl group or a sulfonyl group; and R³ is aC₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group,a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅aralkyl group, a C₇ -C₁₅ aralkylamino group or a heterocyclic groupwhich is not substituted, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which has one to foursubstituents selected from the group consisting of halogen atoms, cyanogroups, hydroxyl groups, amino groups and carboxyl groups, and hydroxylgroups, amino groups and carboxyl groups having a C₁ -C₁₆ alkyl group, ahalo-C₁ -C₁₆ alkyl group, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁-C₁₆ alkyl group, a carboxy-C₁ -C₁₆ alkyl group or a protecting groupprovided that when R² is a hydrogen atom, R¹ is not a methyl group or anacetyl group; when R² is an acetyl group, R¹ is not a methyl group. 2.The compound according to claim 1, which is a compound represented bygeneral formula (I-a): ##STR9## wherein R^(1a) is a hydrogen atom, a C₁-C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆-C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic group whichis not substituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃-C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which has one to five substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, C₁-C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxy groups, amino groups,mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆ alkylamino groups, carboxylgroups, C₁ -C₁₆ alkyloxycarbonyl groups, aminocarbonyl groups, sulfogroups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxy groups, C₇ -C₁₅ aralkyloxygroups and heterocyclic groups, or a group represented by --Y^(a)--R^(3a) ; Y^(a) is a carbonyl group, a thiocarbonyl group or a sulfonylgroup; R^(3a) is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃-C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or aheterocyclic group which is not substituted, or a C₁ -C₁₆ alkyl group, aC₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁-C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇-C₁₅ aralkylamino group or a heterocyclic group which has one to foursubstituents selected from the group consisting of halogen atoms, cyanogroups, hydroxyl groups, amino groups and carboxyl groups, and hydroxylgroups, amino groups and carboxyl groups having a C₁ -C₁₆ alkyl group, ahalo-C₁ -C₁₆ alkyl group, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁-C₁₆ alkyl group, a carboxy-C₁ -C₁₆ alkyl group or a protecting group;R^(2a) is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which is not substituted, a C₁ -C₁₆ alkyl group, a C₂-C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇-C₁₅ aralkyl group or a heterocyclic group which has one to fivesubstituents selected from the group consisting of halogen atoms, cyanogroups, hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆alkylcarbonyloxy groups, amino groups, mono-C₁ -C₁₆ alkylamino groups,di-C₁ -C₁₆ alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonylgroups, aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂aryloxy groups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or agroup represented by --Y^(b) --R^(3b) ; Y^(b) is a carbonyl group, athiocarbonyl group or a sulfonyl group; and R^(3b) is a C₁ -C₁₆ alkylgroup, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup, a C₇ -C₁₅ aralkylamino group or a heterocyclic group which is notsubstituted, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or aheterocyclic group which has one to four substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, aminogroups and carboxyl groups, and hydroxyl groups, amino groups andcarboxyl groups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkylgroup, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, acarboxy-C₁ -C₁₆ alkyl group or a protecting group provided that whenY^(b) is a carbonyl group, and R^(3b) is a C₁ -C₁₆ alkyl group which isnot substituted, R^(1a) is not a C₁ -C₁₆ alkyl group which is notsubstituted.
 3. The compound according to claim 1, which is a compoundrepresented by general formula (I-b): ##STR10## wherein R^(1b) is a C₁-C₁₆ alkyl group which is not substituted; R^(2b) is a group representedby --Y^(c) --R^(3c) ; Y^(c) is a carbonyl group; and R^(3c) is a C₁ -C₁₆alkyl group which is not substituted provided that both R^(1b) andR^(3c) are not methyl groups at the same time.
 4. The compound accordingto claim 1, which is a compound represented by general formula (I-c):##STR11## wherein R^(1c) is a hydrogen atom, a C₂ -C₁₀ alkenyl group, aC₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, aC₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or aheterocyclic group which has one to five substituents selected from thegroup consisting of halogen atoms, cyano groups, hydroxyl groups, C₁-C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxy groups, amino groups,mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆ alkylamino groups, carboxylgroups, C₁ -C₁₆ alkyloxycarbonyl groups, aminocarbonyl groups, sulfogroups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxy groups, C₇ -C₁₅ aralkyloxygroups and heterocyclic groups, or a group represented by --Y--R³ ; Y isa carbonyl group, a thiocarbonyl group or a sulfonyl group; R³ is a C₁-C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkyl group, aC₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅aralkylamino group, a C₇ -C₁₅ aralkylamino group or a heterocyclic groupwhich is not substituted, or a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which has one to foursubstituents selected from the group consisting of halogen atoms, cyanogroups, hydroxyl groups, amino groups and carboxyl groups, and hydroxylgroups, amino groups and carboxyl groups having a C₁ -C₁₆ alkyl group, ahalo-C₁ -C₁₆ alkyl group, a hydroxy-C₁ -C₁₆ alkyl group, an amino-C₁-C₁₆ alkyl group, a carboxy-C₁ -C₁₆ alkyl group or a protecting group;and R^(2c) is a hydrogen atom provided that when Y is a carbonyl group,R³ is not a C₁ -C₁₆ alkyl group which is not substituted.
 5. Thecompound according to claim 1, which is a compound represented bygeneral formula (I-d): ##STR12## wherein R^(1d) is a C₁ -C₁₆ alkyl groupwhich is not substituted; and R^(2c) is a hydrogen atom provided thatR^(1d) is not a methyl group.
 6. The compound according to claim 5,wherein R^(1d) is a C₄ -C₁₀ alkyl group which is not substituted.
 7. Thecompound according to claim 5, wherein R^(1d) is a butyl group, a pentylgroup, an isopentyl group, a hexyl group or a decyl group.
 8. Thecompound according to claim 1, which is a compound represented bygeneral formula (I-e): ##STR13## wherein R^(1e) is a group representedby --Y^(c) --R^(3d) ; Y^(c) is a carbonyl group; R^(3d) is a C₁ -C₁₆alkyl group which is not substituted; and R^(2c) is a hydrogen atomprovided that R^(3d) is not a methyl group.
 9. The compound according toclaim 8, wherein R^(3d) is a C₃ -C₉ alkyl group which is notsubstituted.
 10. The compound according to claim 8, wherein R^(3d) is apropyl group, a butyl group, a pentyl group, a hexyl group or a nonylgroup.
 11. An antifungal agent containing a compound represented bygeneral formula (I) or a pharmaceutically acceptable salt or esterthereof: ##STR14## wherein each of R¹ and R² is independently a hydrogenatom, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclicgroup which is not substituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup or a heterocyclic group which has one to five substituentsselected from the group consisting of halogen atoms, cyano groups,hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxygroups, amino groups, mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonyl groups,aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxygroups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or a grouprepresented by --Y--R³ ; Y is a carbonyl group, a thiocarbonyl group ora sulfonyl group; and R³ is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which is not substituted, ora C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkylgroup, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, aC₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or a heterocyclicgroup which has one to four substituents selected from the groupconsisting of halogen atoms, cyano groups, hydroxyl groups, amino groupsand carboxyl groups, and hydroxyl groups, amino groups and carboxylgroups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkyl group, ahydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, a carboxy-C₁-C₁₆ alkyl group or a protecting group, provided that when R² is ahydrogen atom, R¹ is not a methyl group or an acetyl group; when R² isan acetyl group, R¹ is not a methyl group, as an active ingredient. 12.An antifungal composition containing a compound represented by generalformula (I) or a pharmaceutically acceptable salt or ester thereof:##STR15## wherein each of R¹ and R² is independently a hydrogen atom, aC₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, aC₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group or a heterocyclic groupwhich is not substituted, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup or a heterocyclic group which has one to five substituentsselected from the group consisting of halogen atoms, cyano groups,hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆ alkylcarbonyloxygroups, amino groups, mono-C₁ -C₁₆ alkylamino groups, di-C₁ -C₁₆alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonyl groups,aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂ aryloxygroups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or a grouprepresented by --Y--R³ ; Y is a carbonyl group, a thiocarbonyl group ora sulfonyl group; and R³ is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkylgroup, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which is not substituted, ora C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkylgroup, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, aC₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or a heterocyclicgroup which has one to four substituents selected from the groupconsisting of halogen atoms, cyano groups, hydroxyl groups, amino groupsand carboxyl groups, and hydroxyl groups, amino groups and carboxylgroups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkyl group, ahydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, a carboxy-C₁-C₁₆ alkyl group or a protecting group, provided that when R² is ahydrogen atom, R¹ is not a methyl group or an acetyl group; when R² isan acetyl group, R¹ is not a methyl group, and an azole type antifungalagent, as active ingredients.
 13. The antifungal composition accordingto claim 12, wherein the azole type antifungal agent is butoconazole,oxiconazole, clotrimazole, terconazole, econazole, tioconazole,miconazole, fluconazole, ketoconazole or itraconazole.
 14. Theantifungal composition according to claim 12, wherein the weight ratioof the compound represented by general formula (I) or a pharmaceuticallyacceptable salt or ester thereof: ##STR16## wherein each of R¹ and R² isindependently a hydrogen atom, a C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenylgroup, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkylgroup or a heterocyclic group which is not substituted, a C₁ -C₁₆ alkylgroup, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ alkynyl group, a C₆ -C₁₂ arylgroup, a C₇ -C₁₅ aralkyl group or a heterocyclic group which has one tofive substituents selected from the group consisting of halogen atoms,cyano groups, hydroxyl groups, C₁ -C₁₆ alkyloxy groups, C₁ -C₁₆alkylcarbonyloxy groups, amino groups, mono-C₁ -C₁₆ alkylamino groups,di-C₁ -C₁₆ alkylamino groups, carboxyl groups, C₁ -C₁₆ alkyloxycarbonylgroups, aminocarbonyl groups, sulfo groups, C₆ -C₁₂ aryl groups, C₆ -C₁₂aryloxy groups, C₇ -C₁₅ aralkyloxy groups and heterocyclic groups, or agroup represented by --Y--R³ ; Y is a carbonyl group, a thiocarbonylgroup or a sulfonyl group; and R³ is a C₁ -C₁₆ alkyl group, a C₂ -C₁₀alkenyl group, a C₃ -C₆ cycloalkyl group, a C₃ -C₆ cycloalkyl-C₁ -C₁₆alkyl group, a C₆ -C₁₂ aryl group, a C₇ -C₁₅ aralkyl group, a C₇ -C₁₅aralkylamino group or a heterocyclic group which is not substituted, ora C₁ -C₁₆ alkyl group, a C₂ -C₁₀ alkenyl group, a C₃ -C₆ cycloalkylgroup, a C₃ -C₆ cycloalkyl-C₁ -C₁₆ alkyl group, a C₆ -C₁₂ aryl group, aC₇ -C₁₅ aralkyl group, a C₇ -C₁₅ aralkylamino group or a heterocyclicgroup which has one to four substituents selected from the groupconsisting of halogen atoms, cyano groups, hydroxyl groups, amino groupsand carboxyl groups, and hydroxyl groups, amino groups and carboxylgroups having a C₁ -C₁₆ alkyl group, a halo-C₁ -C₁₆ alkyl group, ahydroxy-C₁ -C₁₆ alkyl group, an amino-C₁ -C₁₆ alkyl group, a carboxy-C₁-C₁₆ alkyl group or a protecting group, provided that when R² is ahydrogen atom, R¹ is not a methyl group or an acetyl group; when R² isan acetyl group, R¹ is not a methyl group, to the azole type antifungalagent is from 0.001:1 to 1000:1.